Through the introduction of disaster situation of Qiang Culture after Wenchuan Earthquake, the paper emphasized that carriers of Qiang Culture had been seriously damaged, the inheritance of Qiang Culture had been affe...Through the introduction of disaster situation of Qiang Culture after Wenchuan Earthquake, the paper emphasized that carriers of Qiang Culture had been seriously damaged, the inheritance of Qiang Culture had been affected, and the environment for Qiang Culture was difficult to recover. It highlighted that three-dimensional reconstruction of Qiang Culture should stress the core task and timely and effectively rescue endangered cultural heritages of Qiang Nationality from the perspectives of material and spiritual life. It had explained focuses of three-dimensional pattern construction in detail. In terms of spatial reconstruction, it should reconstruct native culture and history while material culture was constructed, and reconstruct Qiang culture highland by depending on aborigines; in terms of cluster reconstruction, it should give support to large tourism enterprises and perfect tourism chain; in terms of ecological reconstruction, it should enhance construction and demonstration of "ecological protection pilot area of Qiang culture"; in terms of development reconstruction, it should realize coordinated unity between protection and development according to classification protection, characteristic protection and key protection, so as to form the virtuous circle of post-disaster recovery protection and sustainable development.展开更多
The cancer stem cells(CSCs)from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified.The primary cells derived from human osteosarcoma were digested by...The cancer stem cells(CSCs)from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified.The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension,and mixed homogeneously into 1.2% alginate gel.Single-cell alginate gel was cultured with serum-free DMEM/F12 medium.Epirubicin(0.8μg/mL)was added to the medium to enrich CSCs.After cultured conventionally for 7 to 10 days,most of cells suspended in ...展开更多
AIM: TO investigate the multicellular resistance of human hepatocellular carcinoma HepG2 cells in three-dimensional culture to delisheng, 5-fluorouracil and adriamycin, and the possible molecular mechanisms of delish...AIM: TO investigate the multicellular resistance of human hepatocellular carcinoma HepG2 cells in three-dimensional culture to delisheng, 5-fluorouracil and adriamycin, and the possible molecular mechanisms of delisheng. METHODS: Human hepatocellular carcinoma HepG2 cells were cultured with a liquid overlay technique. After the formation of multicellular spheroids, morphology was analyzed by phase contrast microscopy, scanning electron microscopy and transmission electron microscopy. Sensitivity of HepG2 cells to delisheng, 5-fluorouracil and adriamycin was investigated by Ml-I- assay in multicelluar spheroids and monolayers. Vascular endothelial growth factor (VEGF) and endostatin expression were analyzed in multicellular spheroids treated with delisheng, 5-fluorouracil, adriamycin and negative control PBS, with immunohistochemical staining. RESULTS: Multicellular spheroids exhibited structural characteristics somewhat different to those in monolayers. The cells in three-dimensional cell culture turned out to be less sensitive to delisheng, 5-fluorouracil and adriamycin than the cells cultured in monolayer. This showed that delisheng had a satisfactory cells inhibition ratio compared to 5-fluorouracil and adriamycin. Immunohistochemical staining showed that VEGF and endostatin expression was positive during growth as multicellular spheroids, and endostatin expression in spheroids with treatment of delisheng was higher than that with 5-fluorouracil, adriamycin and PBS (139.35 ± 7.83, 159.23 ± 10.34, 162.83 ± 3.47 and 148.48 ± 11.06, P 〈 0.05).CONCLUSION: Chinese medicine compound delisheng has satisfactory anti-tumor activity in HepG2 cells in three-dimensional culture, and the effects are associated with up-regulation of endostatin.展开更多
Three-dimensional(3D)culture systems are becoming increasingly popular due to their ability to mimic tissue-like structures more effectively than the monolayer cultures.In cancer and stem cell research,the natural cel...Three-dimensional(3D)culture systems are becoming increasingly popular due to their ability to mimic tissue-like structures more effectively than the monolayer cultures.In cancer and stem cell research,the natural cell characteristics and architectures are closely mimicked by the 3D cell models.Thus,the 3D cell cultures are promising and suitable systems for various proposes,ranging from disease modeling to drug target identification as well as potential therapeutic substances that may transform our lives.This review provides a comprehensive compendium of recent advancements in culturing cells,in particular cancer and stem cells,using 3D culture techniques.The major approaches highlighted here include cell spheroids,hydrogel embedding,bioreactors,scaffolds,and bioprinting.In addition,the progress of employing 3D cell culture systems as a platform for cancer and stem cell research was addressed,and the prominent studies of 3D cell culture systems were discussed.展开更多
AIM: To devise a simplified and efficient method for long-term culture and maintenance of embryonic stem cells requiring less frequent passaging. METHODS: Mouse embryonic stem cells(ESCs) labeled with enhanced yellow ...AIM: To devise a simplified and efficient method for long-term culture and maintenance of embryonic stem cells requiring less frequent passaging. METHODS: Mouse embryonic stem cells(ESCs) labeled with enhanced yellow fluorescent protein were cultured in three-dimensional(3-D) self-assembling scaffolds and compared with traditional two-dimentional(2-D) culture techniques requiring mouse embryonic fibroblast feeder layers or leukemia inhibitory factor. 3-D scaffolds encapsulating ESCs were prepared by mixing ESCs with polyethylene glycol tetra-acrylate(PEG-4-Acr) and thiolfunctionalized dextran(Dex-SH). Distribution of ESCs in 3-D was monitored by confocal microscopy. Viability and proliferation of encapsulated cells during long-term culture were determined by propidium iodide as well as direct cell counts and PrestoB lue(PB) assays. Genetic expression of pluripotency markers(Oct4, Nanog, Klf4, and Sox2) in ESCs grown under 2-D and 3-D cultureconditions was examined by quantitative real-time polymerase chain reaction. Protein expression of selected stemness markers was determined by two different methods, immunofluorescence staining(Oct4 and Nanog) and western blot analysis(Oct4, Nanog, and Klf4). Pluripotency of 3-D scaffold grown ESCs was analyzed by in vivo teratoma assay and in vitro differentiation via embryoid bodies into cells of all three germ layers. RESULTS: Self-assembling scaffolds encapsulating ESCs for 3-D culture without the loss of cell viability were prepared by mixing PEG-4-Acr and Dex-SH(1:1 v/v) to a final concentration of 5%(w/v). Scaffold integrity was dependent on the degree of thiol substitution of Dex-SH and cell concentration. Scaffolds prepared using Dex-SH with 7.5% and 33% thiol substitution and incubated in culture medium maintained their integrity for 11 and 13 d without cells and 22 ± 5 d and 37 ± 5 d with cells, respectively. ESCs formed compact colonies, which progressively increased in size over time due to cell proliferation as determined by confocal microscopy and PB staining. 3-D scaffold cultured ESCs expressed significantly higher levels(P < 0.01) of Oct4, Nanog, and Kl4, showing a 2.8, 3.0 and 1.8 fold increase, respectively, in comparison to 2-D grown cells. A similar increase in the protein expression levels of Oct4, Nanog, and Klf4 was observed in 3-D grown ESCs. However, when 3-D cultured ESCs were subsequently passaged in 2-D culture conditions, the level of these pluripotent markers was reduced to normal levels. 3-D grown ESCs produced teratomas and yielded cells of all three germ layers, expressing brachyury(mesoderm), NCAM(ectoderm), and GATA4(endoderm) markers. Furthermore, these cells differentiated into osteogenic, chondrogenic, myogenic, and neural lineages expressing Col1, Col2, Myog, and Nestin, respectively. CONCLUSION: This novel 3-D culture system demonstrated long-term maintenance of mouse ESCs without the routine passaging and manipulation necessary for traditional 2-D cell propagation.展开更多
Although the recent advances in stem cell engineering have gained a great deal of attention due to their high potential in clinical research,the applicability of stem cells for preclinical screening in the drug discov...Although the recent advances in stem cell engineering have gained a great deal of attention due to their high potential in clinical research,the applicability of stem cells for preclinical screening in the drug discovery process is still challenging due to difficulties in controlling the stem cell microenvironment and the limited availability of high-throughput systems.Recently,researchers have been actively developing and evaluating three-dimensional(3D)cell culture-based platforms using microfluidic technologies,such as organ-on-a-chip and organoid-on-a-chip platforms,and they have achieved promising breakthroughs in stem cell engineering.In this review,we start with a comprehensive discussion on the importance of microfluidic 3D cell culture techniques in stem cell research and their technical strategies in the field of drug discovery.In a subsequent section,we discuss microfluidic 3D cell culture techniques for high-throughput analysis for use in stem cell research.In addition,some potential and practical applications of organ-on-a-chip or organoid-on-a-chip platforms using stem cells as drug screening and disease models are highlighted.展开更多
Objective This study optimizes three-dimensional(3D) culture conditions of HepG2 using response surface methodology(RSM) based on the VitroGel system to facilitate the cell model in vitro for liver tissues.Method HepG...Objective This study optimizes three-dimensional(3D) culture conditions of HepG2 using response surface methodology(RSM) based on the VitroGel system to facilitate the cell model in vitro for liver tissues.Method HepG2 cell was 3D cultured on the VitroGel system.Cell viability was detected using Cell Counting Kit-8(CCK-8) assay of HepG2 lived cell numbers.The proliferation of HepG2 cell and clustering performance was measured via fluorescence staining test.Albumin concentration in the culture medium supernatant as an index of HepG2 cell biological function was measured with ELISA kit.Independent factor tests were conducted with three key factors:inoculated cell concentration,cultured time,and dilution degree of the hydrogel.The preliminary results of independent factor tests were used to determine the levels of factors for RSM.Result The selected optimal culture conditions are as follows:concentration of inoculated cells was4.44 × 10^(5)/mL,culture time was 4.86 days,and hydrogel dilution degree was 1:2.23.The result shows that under optimal conditions,the predicted optical density(OD) value of cell viability was 3.10 and measured 2.978 with a relative error of 3.94%.Conclusion This study serves as a reference for the 3D HepG2 culture and constructs liver tissues in vitro.Additionally,it provides the foundation for repeated dose high-throughput toxicity studies and other scientific research work.展开更多
Objective Adipose tissue distributes widely in human body. The irradiation response of the adipose cells in vivo remains to be investigated. In this study we investigated irradiation response of adipose-derived stem c...Objective Adipose tissue distributes widely in human body. The irradiation response of the adipose cells in vivo remains to be investigated. In this study we investigated irradiation response of adipose-derived stem cells (ASCs) under three-dimensional culture condition. Methods ASCs were isolated and cultured in low attachment dishes to form three-dimensional (3D) spheres in vitro. The neuronal differentiation potential and stem-liked characteristics was monitored by using immunofluoresence staining and flow cytometry in monolayer and 3D culture. To investigate the irradiation sensitivity of 3D sphere culture, the fraction of colony survival and micronucleus were detected in monolayer and 3D culture. Soft agar assays were performed for measuring malignant transformation for the irradiated monolayer and 3D culture. Results The 3D cultured ASCs had higher differentiation potential and an higher stem-like cell percentage. The 3D cultures were more radioresistant after either high linear energy transfer (LET) carbon ion beam or low LET X-ray irradiation compared with the monolayer cell. The ASCs’ potential of cellular transformation was lower after irradiation by soft agar assay. Conclusion These findings suggest that adipose tissue cell are relatively genomic stable and resistant to genotoxic stress.展开更多
Inhibition of mammalian target of rapamycin (m- TOR) is a potential method for cancer treatment. Effects of rapamycin (RAP) on the reversion of malignant breast epithelial cells were investigated on three-dimensional ...Inhibition of mammalian target of rapamycin (m- TOR) is a potential method for cancer treatment. Effects of rapamycin (RAP) on the reversion of malignant breast epithelial cells were investigated on three-dimensional (3D) basement membrane extract (BME) cultures. Through continuous exposure to 20 nM of RAP, cell colony size was significantly reduced in 3D BME cultures of malignant breast epithelial cells, while normal cell colony size appeared unaffected. In unfixed 3D BME cultures of normal and RAP-treated malignant breast epithelial cells, the presence of luminal cell death was confirmed by ethidium bromide and propidium iodide labeling. Increased structural organization was observed by im- munofluorescence staining of F-actin and β-catenin in RAP-treated malignant breast epithelial cells. In monolayer cultures of normal and malignant breast epithelial cells, continuous exposure to 20 nM of RAP increased caspase 3/7 activity and decreased proliferation. Reverse transcriptase polymerase ch- ain reaction (RT-PCR) array analysis indicated a fold increase in the expression of a number of proteins related to polarity, cell-cell adhesion, and cell-matrix adhesion in the presence of RAP. Our data showed that phenotypic reversion of malignancy can be ach- ieved through RAP exposure on 3D BME cultures. This 3D BME culture system will provide correct microenvironments for observing the effects of other mTOR inhibitors on phenotypic reversion of malignant breast epithelial cells.展开更多
In order to obtain a better sandstone three-dimensional (3D) reconstruction result which is more similar to the original sample, an algorithm based on stationarity for a two-dimensional (2D) training image is prop...In order to obtain a better sandstone three-dimensional (3D) reconstruction result which is more similar to the original sample, an algorithm based on stationarity for a two-dimensional (2D) training image is proposed. The second-order statistics based on texture features are analyzed to evaluate the scale stationarity of the training image. The multiple-point statistics of the training image are applied to obtain the multiple-point statistics stationarity estimation by the multi-point density function. The results show that the reconstructed 3D structures are closer to reality when the training image has better scale stationarity and multiple-point statistics stationarity by the indications of local percolation probability and two-point probability. Moreover, training images with higher multiple-point statistics stationarity and lower scale stationarity are likely to obtain closer results to the real 3D structure, and vice versa. Thus, stationarity analysis of the training image has far-reaching significance in choosing a better 2D thin section image for the 3D reconstruction of porous media. Especially, high-order statistics perform better than low-order statistics.展开更多
With the development of globalization and gradual intensification of mutual dependence of the world, "hard power" which includes economic and military strength is no longer the primary consideration for many...With the development of globalization and gradual intensification of mutual dependence of the world, "hard power" which includes economic and military strength is no longer the primary consideration for many countries. They tend to pay more attention to the attractiveness of their "soft power",which includes values,life style , ideology and so forth. Just as Joseph S. Ny , the famous professor of Harvard who puts forward the concept of soft power, says ,"only by widespread communication and diffusion can a country reinforces its soft power". This is mass media's function. Since the 1990s, tens of thousands cultural products brought in from America are exported to foreign countries, including popular music, films ,TV programs, magazines, books and so forth. By making advantage of its monopoly position in media circle, the United States pursues cultural hegemonism and expands its popular culture all over the world. This thesis takes a general view of the great advantages of America multi-national media groups and how they push forward the American popular culture all over the world.展开更多
In the process of meida Convergence,many researchers paid excessive attention to media technology, industry and management,and ignored the culture dimensions of media convergence. Therefore,to transcend media converge...In the process of meida Convergence,many researchers paid excessive attention to media technology, industry and management,and ignored the culture dimensions of media convergence. Therefore,to transcend media convergence technology, industrial thinking and more to the particularity attach importance to cultural media, it is a right way to achieve media convergence. But in the context of China's culture,media convergence should value the cultural uniqueness and the imbalance of the realistic problems,to reach innovation and breakthrough.展开更多
Objective This study aimed to investigate the influence of different culture media on early embryonic cleavage kinetics using time-lapse analysis and to determine the possible relationships between energy substrates i...Objective This study aimed to investigate the influence of different culture media on early embryonic cleavage kinetics using time-lapse analysis and to determine the possible relationships between energy substrates in culture media and the cleavage kinetics.Methods A total of 10021 embryos from 1310 couples were cultured in time-lapse incubators.Embryos cultured in Vitrolife media were allocated to group I,and those in COOK media to group II.Embryo cleavage time points up to the 8-cell stage(t2–t8)were observed after pronuclei fading.Results The baseline demographic features,in vitro fertilization indications,ovarian stimulation protocol,oocyte-cumulus complexes,fertilization rate,together with pregnancy and perinatal outcomes were similar(P>0.05)between groups I and II.According to the time-lapse analysis,all embryos in group I showed significantly faster cleavage speed than those in group II(P<0.05).Furthermore,there was better synchrony in division(s3)and a longer length of the third cell cycle duration(cc3)in group II.Interestingly,implanted embryos in group II showed faster cleavage speed than those in group I,especially at t4 and t7.The glucose contents and multiple major amino acids were similar between the two groups.Lactic and pyruvic acid contents were generally higher in group I than those in group II.Conclusion Because different commercial culture media may influence cleavage kinetics of embryos,it is essential for embryologists to take culture media into consideration in selecting a potential embryo when using a time-lapse system before implantation.展开更多
Aim To develop a sensitive competitive ELBA for the determination of biotinin transformed yeast culture media. Methods The ELBA plate was firstly coated with Mycoplasmahyopneumoniae, and then successively incubated wi...Aim To develop a sensitive competitive ELBA for the determination of biotinin transformed yeast culture media. Methods The ELBA plate was firstly coated with Mycoplasmahyopneumoniae, and then successively incubated with rabbit anti-Mycoplasma hyopneumoniae serum andgoat anti-rabbit IgG-biotin to form the solid biotin, which competed with the biotin in the solution(standard or sample) for the limited streptavidin-horse radish peroxidase conjugate. The standardcalibration curve for biotin analysis was constructed in the range of 50 - 2000 ng·L^(-1). ResultsThe detection limit for biotin was found to be 83 ng·L^(-1), which was about 1000 times lower thanthe lowest determination concentration in the reported ELISA for biotin analysis. The relativestandard deviations for the spiked samples at biotin concentrations of 200 ng·L^(-1), 500ng·L^(-1), and 1000 ng·L^(-1) were 24.87%, 6.15% , and 7.86% , respectively, with the averagerecovery of 101.13% . The wild yeast and its sixty-three transformed yeast culture media wereapplied to the developed ELBA for the determination of biotin. It was found that the biotinconcentrations in more than 85% of the tested samples were enhanced with different increase factorsafter transformation. Conclusion Utilization of Mycoplasma hyopneumoniae as the coating proteinimproves the precision and accuracy of the ELBA assay, which might be used for the biotin assay inother media.展开更多
By analyzing the dilemma encountered in practical teaching of new media advertising course in the digital economy era, this paper explores the teaching path of new media advertising course based on “three-dimensional...By analyzing the dilemma encountered in practical teaching of new media advertising course in the digital economy era, this paper explores the teaching path of new media advertising course based on “three-dimensional” practical teaching system, and believes that it is necessary to accurately determine talent training objectives, and construct the three dimensions of practical teaching, practical ability, and new media entrepreneurship and employment. Finally, the teaching countermeasures of practical teaching system based on three dimensions are put forward.展开更多
Neuronal cell death and the loss of connectivity are two of the primary pathological mechanisms underlying Alzheimer's disease.The accumulation of amyloid-βpeptides,a key hallmark of Alzheimer's disease,is be...Neuronal cell death and the loss of connectivity are two of the primary pathological mechanisms underlying Alzheimer's disease.The accumulation of amyloid-βpeptides,a key hallmark of Alzheimer's disease,is believed to induce neuritic abnormalities,including reduced growth,extension,and abnormal growth cone morphology,all of which contribute to decreased connectivity.However,the precise cellular and molecular mechanisms governing this response remain unknown.In this study,we used an innovative approach to demonstrate the effect of amyloid-βon neurite dynamics in both two-dimensional and three-dimensional cultu re systems,in order to provide more physiologically relevant culture geometry.We utilized various methodologies,including the addition of exogenous amyloid-βpeptides to the culture medium,growth substrate coating,and the utilization of human-induced pluripotent stem cell technology,to investigate the effect of endogenous amyloid-βsecretion on neurite outgrowth,thus paving the way for potential future applications in personalized medicine.Additionally,we also explore the involvement of the Nogo signaling cascade in amyloid-β-induced neurite inhibition.We demonstrate that inhibition of downstream ROCK and RhoA components of the Nogo signaling pathway,achieved through modulation with Y-27632(a ROCK inhibitor)and Ibuprofen(a Rho A inhibitor),respectively,can restore and even enhance neuronal connectivity in the presence of amyloid-β.In summary,this study not only presents a novel culture approach that offers insights into the biological process of neurite growth and inhibition,but also proposes a specific mechanism for reduced neural connectivity in the presence of amyloid-βpeptides,along with potential intervention points to restore neurite growth.Thereby,we aim to establish a culture system that has the potential to serve as an assay for measuring preclinical,predictive outcomes of drugs and their ability to promote neurite outgrowth,both generally and in a patient-specific manner.展开更多
With the rise of new media and short videos shaping the new communication environment,rural culture has been mediated and transformed to spread to a wider region.As a cultural achievement of rural revitalization,“Vil...With the rise of new media and short videos shaping the new communication environment,rural culture has been mediated and transformed to spread to a wider region.As a cultural achievement of rural revitalization,“Village BA”(Village Basketball Association)demonstrates Chinese modernization.The use of short videos,mass rural movements,and“spectacle”spaces to attract spectators has become a key issue in the dissemination of rural culture.展开更多
The three-dimensional(3D)cell culture system has garnered significant attention in recent years as a means of studying cell behavior and tissue development,as opposed to traditional two-dimensional cultures.These syst...The three-dimensional(3D)cell culture system has garnered significant attention in recent years as a means of studying cell behavior and tissue development,as opposed to traditional two-dimensional cultures.These systems can induce specific cell reactions,promote specific tissue functions,and serve as valuable tools for research in tissue engineering,regenerative medicine,and drug discovery.This paper discusses current developments in the field of three-dimensional cell culture and the potential applications of 3D type 1 collagen gels to enhance the growth and maturation of dendritic cells.展开更多
In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were in...In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were induced to form callus when cultured on vermicompost extract media along with coelomic fluid. Suspension medium was developed using vermicompost extract and coelomic fluid in 3:1 ratio. Phytochemical analysis of the alkaloid berberine was confirmed from callus, suspension cell culture and suspension medium by Thin Layer Chromatography and High Performance Liquid Chromatography. Vermicompost and its extracts with coelomic fluid have shown maximum (100 per cent) response of callus induction. Callus mass enlarged with increasing concentration of coelomic fluid and callus growth was assessed from the biomass. Incubation of culture tubes in dark supported callus development significantly. The Rf value of 0.36 confirmed the presence of berberine by Thin Layer Chromatography. Qualitative analysis confirmed the presence of alkaloid berberine with the retention time of 2.8 minutes similar to that of standard reference sample from Sigma chemicals, USA. The suspension medium turned deep yellow because of the release of the alkaloid. Vermicompost and its extracts along with coelomic fluid have shown the economical approach for micropropagation of economically and medicinally important plants.展开更多
Nowadays,the communication of traditional culture is facing various challenges,including limitations of traditional culture itself,decrease of inheritors,poor construction of structure,and heterization of connotation,...Nowadays,the communication of traditional culture is facing various challenges,including limitations of traditional culture itself,decrease of inheritors,poor construction of structure,and heterization of connotation,as well as conflicts between Chinese and foreign cultures in the context of globalization.New media has provided more opportunities for the communication of outstanding Chinese traditional culture.This paper analyzed the role of new media in completing the structure and expanding the scope of communication of outstanding Chinese traditional culture,and proposed strategy for optimizing the communication route from the perspective of improving environment,changing content,and adjusting strategy.The contributions of new media to the communication of Chinese traditional culture should be highly appreciated.展开更多
文摘Through the introduction of disaster situation of Qiang Culture after Wenchuan Earthquake, the paper emphasized that carriers of Qiang Culture had been seriously damaged, the inheritance of Qiang Culture had been affected, and the environment for Qiang Culture was difficult to recover. It highlighted that three-dimensional reconstruction of Qiang Culture should stress the core task and timely and effectively rescue endangered cultural heritages of Qiang Nationality from the perspectives of material and spiritual life. It had explained focuses of three-dimensional pattern construction in detail. In terms of spatial reconstruction, it should reconstruct native culture and history while material culture was constructed, and reconstruct Qiang culture highland by depending on aborigines; in terms of cluster reconstruction, it should give support to large tourism enterprises and perfect tourism chain; in terms of ecological reconstruction, it should enhance construction and demonstration of "ecological protection pilot area of Qiang culture"; in terms of development reconstruction, it should realize coordinated unity between protection and development according to classification protection, characteristic protection and key protection, so as to form the virtuous circle of post-disaster recovery protection and sustainable development.
文摘The cancer stem cells(CSCs)from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified.The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension,and mixed homogeneously into 1.2% alginate gel.Single-cell alginate gel was cultured with serum-free DMEM/F12 medium.Epirubicin(0.8μg/mL)was added to the medium to enrich CSCs.After cultured conventionally for 7 to 10 days,most of cells suspended in ...
文摘AIM: TO investigate the multicellular resistance of human hepatocellular carcinoma HepG2 cells in three-dimensional culture to delisheng, 5-fluorouracil and adriamycin, and the possible molecular mechanisms of delisheng. METHODS: Human hepatocellular carcinoma HepG2 cells were cultured with a liquid overlay technique. After the formation of multicellular spheroids, morphology was analyzed by phase contrast microscopy, scanning electron microscopy and transmission electron microscopy. Sensitivity of HepG2 cells to delisheng, 5-fluorouracil and adriamycin was investigated by Ml-I- assay in multicelluar spheroids and monolayers. Vascular endothelial growth factor (VEGF) and endostatin expression were analyzed in multicellular spheroids treated with delisheng, 5-fluorouracil, adriamycin and negative control PBS, with immunohistochemical staining. RESULTS: Multicellular spheroids exhibited structural characteristics somewhat different to those in monolayers. The cells in three-dimensional cell culture turned out to be less sensitive to delisheng, 5-fluorouracil and adriamycin than the cells cultured in monolayer. This showed that delisheng had a satisfactory cells inhibition ratio compared to 5-fluorouracil and adriamycin. Immunohistochemical staining showed that VEGF and endostatin expression was positive during growth as multicellular spheroids, and endostatin expression in spheroids with treatment of delisheng was higher than that with 5-fluorouracil, adriamycin and PBS (139.35 ± 7.83, 159.23 ± 10.34, 162.83 ± 3.47 and 148.48 ± 11.06, P 〈 0.05).CONCLUSION: Chinese medicine compound delisheng has satisfactory anti-tumor activity in HepG2 cells in three-dimensional culture, and the effects are associated with up-regulation of endostatin.
文摘Three-dimensional(3D)culture systems are becoming increasingly popular due to their ability to mimic tissue-like structures more effectively than the monolayer cultures.In cancer and stem cell research,the natural cell characteristics and architectures are closely mimicked by the 3D cell models.Thus,the 3D cell cultures are promising and suitable systems for various proposes,ranging from disease modeling to drug target identification as well as potential therapeutic substances that may transform our lives.This review provides a comprehensive compendium of recent advancements in culturing cells,in particular cancer and stem cells,using 3D culture techniques.The major approaches highlighted here include cell spheroids,hydrogel embedding,bioreactors,scaffolds,and bioprinting.In addition,the progress of employing 3D cell culture systems as a platform for cancer and stem cell research was addressed,and the prominent studies of 3D cell culture systems were discussed.
基金Oakland University and Oakland University-William Beaumont Institute for Stem Cell and Regenerative Medicine(OU-WB ISCRM)
文摘AIM: To devise a simplified and efficient method for long-term culture and maintenance of embryonic stem cells requiring less frequent passaging. METHODS: Mouse embryonic stem cells(ESCs) labeled with enhanced yellow fluorescent protein were cultured in three-dimensional(3-D) self-assembling scaffolds and compared with traditional two-dimentional(2-D) culture techniques requiring mouse embryonic fibroblast feeder layers or leukemia inhibitory factor. 3-D scaffolds encapsulating ESCs were prepared by mixing ESCs with polyethylene glycol tetra-acrylate(PEG-4-Acr) and thiolfunctionalized dextran(Dex-SH). Distribution of ESCs in 3-D was monitored by confocal microscopy. Viability and proliferation of encapsulated cells during long-term culture were determined by propidium iodide as well as direct cell counts and PrestoB lue(PB) assays. Genetic expression of pluripotency markers(Oct4, Nanog, Klf4, and Sox2) in ESCs grown under 2-D and 3-D cultureconditions was examined by quantitative real-time polymerase chain reaction. Protein expression of selected stemness markers was determined by two different methods, immunofluorescence staining(Oct4 and Nanog) and western blot analysis(Oct4, Nanog, and Klf4). Pluripotency of 3-D scaffold grown ESCs was analyzed by in vivo teratoma assay and in vitro differentiation via embryoid bodies into cells of all three germ layers. RESULTS: Self-assembling scaffolds encapsulating ESCs for 3-D culture without the loss of cell viability were prepared by mixing PEG-4-Acr and Dex-SH(1:1 v/v) to a final concentration of 5%(w/v). Scaffold integrity was dependent on the degree of thiol substitution of Dex-SH and cell concentration. Scaffolds prepared using Dex-SH with 7.5% and 33% thiol substitution and incubated in culture medium maintained their integrity for 11 and 13 d without cells and 22 ± 5 d and 37 ± 5 d with cells, respectively. ESCs formed compact colonies, which progressively increased in size over time due to cell proliferation as determined by confocal microscopy and PB staining. 3-D scaffold cultured ESCs expressed significantly higher levels(P < 0.01) of Oct4, Nanog, and Kl4, showing a 2.8, 3.0 and 1.8 fold increase, respectively, in comparison to 2-D grown cells. A similar increase in the protein expression levels of Oct4, Nanog, and Klf4 was observed in 3-D grown ESCs. However, when 3-D cultured ESCs were subsequently passaged in 2-D culture conditions, the level of these pluripotent markers was reduced to normal levels. 3-D grown ESCs produced teratomas and yielded cells of all three germ layers, expressing brachyury(mesoderm), NCAM(ectoderm), and GATA4(endoderm) markers. Furthermore, these cells differentiated into osteogenic, chondrogenic, myogenic, and neural lineages expressing Col1, Col2, Myog, and Nestin, respectively. CONCLUSION: This novel 3-D culture system demonstrated long-term maintenance of mouse ESCs without the routine passaging and manipulation necessary for traditional 2-D cell propagation.
基金supported by the National Research Foundation of Korea (NRF) (NRF2017R1C1B2002377, NRF-2016R1A5A1010148, and NRF2019R1A2C1003111)funded by the Ministry of Science and ICT (MSIT)partly supported by the Technology Innovation Program (No.10067787)funded by the Ministry of Trade, Industry & Energy (MOTE, Korea)
文摘Although the recent advances in stem cell engineering have gained a great deal of attention due to their high potential in clinical research,the applicability of stem cells for preclinical screening in the drug discovery process is still challenging due to difficulties in controlling the stem cell microenvironment and the limited availability of high-throughput systems.Recently,researchers have been actively developing and evaluating three-dimensional(3D)cell culture-based platforms using microfluidic technologies,such as organ-on-a-chip and organoid-on-a-chip platforms,and they have achieved promising breakthroughs in stem cell engineering.In this review,we start with a comprehensive discussion on the importance of microfluidic 3D cell culture techniques in stem cell research and their technical strategies in the field of drug discovery.In a subsequent section,we discuss microfluidic 3D cell culture techniques for high-throughput analysis for use in stem cell research.In addition,some potential and practical applications of organ-on-a-chip or organoid-on-a-chip platforms using stem cells as drug screening and disease models are highlighted.
基金funded by Toxicity Evaluation of Key Contaminants in Health Food by Cell-based Test Models and the Mechanism Analysis [2018YFC1602104]
文摘Objective This study optimizes three-dimensional(3D) culture conditions of HepG2 using response surface methodology(RSM) based on the VitroGel system to facilitate the cell model in vitro for liver tissues.Method HepG2 cell was 3D cultured on the VitroGel system.Cell viability was detected using Cell Counting Kit-8(CCK-8) assay of HepG2 lived cell numbers.The proliferation of HepG2 cell and clustering performance was measured via fluorescence staining test.Albumin concentration in the culture medium supernatant as an index of HepG2 cell biological function was measured with ELISA kit.Independent factor tests were conducted with three key factors:inoculated cell concentration,cultured time,and dilution degree of the hydrogel.The preliminary results of independent factor tests were used to determine the levels of factors for RSM.Result The selected optimal culture conditions are as follows:concentration of inoculated cells was4.44 × 10^(5)/mL,culture time was 4.86 days,and hydrogel dilution degree was 1:2.23.The result shows that under optimal conditions,the predicted optical density(OD) value of cell viability was 3.10 and measured 2.978 with a relative error of 3.94%.Conclusion This study serves as a reference for the 3D HepG2 culture and constructs liver tissues in vitro.Additionally,it provides the foundation for repeated dose high-throughput toxicity studies and other scientific research work.
基金supported by grants from the National Key Scientific Instrument and Equipment Development Project of China[2012YQ03014210]Major Project Specialized for Infectious Diseases of the Chinese Health and Family Planning Commission[2014ZX10004002-004-002]National Natural Science Foundation of China[31170803]to BH
文摘Objective Adipose tissue distributes widely in human body. The irradiation response of the adipose cells in vivo remains to be investigated. In this study we investigated irradiation response of adipose-derived stem cells (ASCs) under three-dimensional culture condition. Methods ASCs were isolated and cultured in low attachment dishes to form three-dimensional (3D) spheres in vitro. The neuronal differentiation potential and stem-liked characteristics was monitored by using immunofluoresence staining and flow cytometry in monolayer and 3D culture. To investigate the irradiation sensitivity of 3D sphere culture, the fraction of colony survival and micronucleus were detected in monolayer and 3D culture. Soft agar assays were performed for measuring malignant transformation for the irradiated monolayer and 3D culture. Results The 3D cultured ASCs had higher differentiation potential and an higher stem-like cell percentage. The 3D cultures were more radioresistant after either high linear energy transfer (LET) carbon ion beam or low LET X-ray irradiation compared with the monolayer cell. The ASCs’ potential of cellular transformation was lower after irradiation by soft agar assay. Conclusion These findings suggest that adipose tissue cell are relatively genomic stable and resistant to genotoxic stress.
文摘Inhibition of mammalian target of rapamycin (m- TOR) is a potential method for cancer treatment. Effects of rapamycin (RAP) on the reversion of malignant breast epithelial cells were investigated on three-dimensional (3D) basement membrane extract (BME) cultures. Through continuous exposure to 20 nM of RAP, cell colony size was significantly reduced in 3D BME cultures of malignant breast epithelial cells, while normal cell colony size appeared unaffected. In unfixed 3D BME cultures of normal and RAP-treated malignant breast epithelial cells, the presence of luminal cell death was confirmed by ethidium bromide and propidium iodide labeling. Increased structural organization was observed by im- munofluorescence staining of F-actin and β-catenin in RAP-treated malignant breast epithelial cells. In monolayer cultures of normal and malignant breast epithelial cells, continuous exposure to 20 nM of RAP increased caspase 3/7 activity and decreased proliferation. Reverse transcriptase polymerase ch- ain reaction (RT-PCR) array analysis indicated a fold increase in the expression of a number of proteins related to polarity, cell-cell adhesion, and cell-matrix adhesion in the presence of RAP. Our data showed that phenotypic reversion of malignancy can be ach- ieved through RAP exposure on 3D BME cultures. This 3D BME culture system will provide correct microenvironments for observing the effects of other mTOR inhibitors on phenotypic reversion of malignant breast epithelial cells.
基金The National Natural Science Foundation of China(No.60972130)
文摘In order to obtain a better sandstone three-dimensional (3D) reconstruction result which is more similar to the original sample, an algorithm based on stationarity for a two-dimensional (2D) training image is proposed. The second-order statistics based on texture features are analyzed to evaluate the scale stationarity of the training image. The multiple-point statistics of the training image are applied to obtain the multiple-point statistics stationarity estimation by the multi-point density function. The results show that the reconstructed 3D structures are closer to reality when the training image has better scale stationarity and multiple-point statistics stationarity by the indications of local percolation probability and two-point probability. Moreover, training images with higher multiple-point statistics stationarity and lower scale stationarity are likely to obtain closer results to the real 3D structure, and vice versa. Thus, stationarity analysis of the training image has far-reaching significance in choosing a better 2D thin section image for the 3D reconstruction of porous media. Especially, high-order statistics perform better than low-order statistics.
文摘With the development of globalization and gradual intensification of mutual dependence of the world, "hard power" which includes economic and military strength is no longer the primary consideration for many countries. They tend to pay more attention to the attractiveness of their "soft power",which includes values,life style , ideology and so forth. Just as Joseph S. Ny , the famous professor of Harvard who puts forward the concept of soft power, says ,"only by widespread communication and diffusion can a country reinforces its soft power". This is mass media's function. Since the 1990s, tens of thousands cultural products brought in from America are exported to foreign countries, including popular music, films ,TV programs, magazines, books and so forth. By making advantage of its monopoly position in media circle, the United States pursues cultural hegemonism and expands its popular culture all over the world. This thesis takes a general view of the great advantages of America multi-national media groups and how they push forward the American popular culture all over the world.
文摘In the process of meida Convergence,many researchers paid excessive attention to media technology, industry and management,and ignored the culture dimensions of media convergence. Therefore,to transcend media convergence technology, industrial thinking and more to the particularity attach importance to cultural media, it is a right way to achieve media convergence. But in the context of China's culture,media convergence should value the cultural uniqueness and the imbalance of the realistic problems,to reach innovation and breakthrough.
基金funded by the National Natural Science Foundation of China(No.82004017 and No.81801531).
文摘Objective This study aimed to investigate the influence of different culture media on early embryonic cleavage kinetics using time-lapse analysis and to determine the possible relationships between energy substrates in culture media and the cleavage kinetics.Methods A total of 10021 embryos from 1310 couples were cultured in time-lapse incubators.Embryos cultured in Vitrolife media were allocated to group I,and those in COOK media to group II.Embryo cleavage time points up to the 8-cell stage(t2–t8)were observed after pronuclei fading.Results The baseline demographic features,in vitro fertilization indications,ovarian stimulation protocol,oocyte-cumulus complexes,fertilization rate,together with pregnancy and perinatal outcomes were similar(P>0.05)between groups I and II.According to the time-lapse analysis,all embryos in group I showed significantly faster cleavage speed than those in group II(P<0.05).Furthermore,there was better synchrony in division(s3)and a longer length of the third cell cycle duration(cc3)in group II.Interestingly,implanted embryos in group II showed faster cleavage speed than those in group I,especially at t4 and t7.The glucose contents and multiple major amino acids were similar between the two groups.Lactic and pyruvic acid contents were generally higher in group I than those in group II.Conclusion Because different commercial culture media may influence cleavage kinetics of embryos,it is essential for embryologists to take culture media into consideration in selecting a potential embryo when using a time-lapse system before implantation.
文摘Aim To develop a sensitive competitive ELBA for the determination of biotinin transformed yeast culture media. Methods The ELBA plate was firstly coated with Mycoplasmahyopneumoniae, and then successively incubated with rabbit anti-Mycoplasma hyopneumoniae serum andgoat anti-rabbit IgG-biotin to form the solid biotin, which competed with the biotin in the solution(standard or sample) for the limited streptavidin-horse radish peroxidase conjugate. The standardcalibration curve for biotin analysis was constructed in the range of 50 - 2000 ng·L^(-1). ResultsThe detection limit for biotin was found to be 83 ng·L^(-1), which was about 1000 times lower thanthe lowest determination concentration in the reported ELISA for biotin analysis. The relativestandard deviations for the spiked samples at biotin concentrations of 200 ng·L^(-1), 500ng·L^(-1), and 1000 ng·L^(-1) were 24.87%, 6.15% , and 7.86% , respectively, with the averagerecovery of 101.13% . The wild yeast and its sixty-three transformed yeast culture media wereapplied to the developed ELBA for the determination of biotin. It was found that the biotinconcentrations in more than 85% of the tested samples were enhanced with different increase factorsafter transformation. Conclusion Utilization of Mycoplasma hyopneumoniae as the coating proteinimproves the precision and accuracy of the ELBA assay, which might be used for the biotin assay inother media.
基金Sponsored by 2022 “Tourism Industry Economic Teaching Innovation Team” of Taishan University。
文摘By analyzing the dilemma encountered in practical teaching of new media advertising course in the digital economy era, this paper explores the teaching path of new media advertising course based on “three-dimensional” practical teaching system, and believes that it is necessary to accurately determine talent training objectives, and construct the three dimensions of practical teaching, practical ability, and new media entrepreneurship and employment. Finally, the teaching countermeasures of practical teaching system based on three dimensions are put forward.
基金supported by a BBSRC CASE training studentship,No.BB/K011413/1(to KG)。
文摘Neuronal cell death and the loss of connectivity are two of the primary pathological mechanisms underlying Alzheimer's disease.The accumulation of amyloid-βpeptides,a key hallmark of Alzheimer's disease,is believed to induce neuritic abnormalities,including reduced growth,extension,and abnormal growth cone morphology,all of which contribute to decreased connectivity.However,the precise cellular and molecular mechanisms governing this response remain unknown.In this study,we used an innovative approach to demonstrate the effect of amyloid-βon neurite dynamics in both two-dimensional and three-dimensional cultu re systems,in order to provide more physiologically relevant culture geometry.We utilized various methodologies,including the addition of exogenous amyloid-βpeptides to the culture medium,growth substrate coating,and the utilization of human-induced pluripotent stem cell technology,to investigate the effect of endogenous amyloid-βsecretion on neurite outgrowth,thus paving the way for potential future applications in personalized medicine.Additionally,we also explore the involvement of the Nogo signaling cascade in amyloid-β-induced neurite inhibition.We demonstrate that inhibition of downstream ROCK and RhoA components of the Nogo signaling pathway,achieved through modulation with Y-27632(a ROCK inhibitor)and Ibuprofen(a Rho A inhibitor),respectively,can restore and even enhance neuronal connectivity in the presence of amyloid-β.In summary,this study not only presents a novel culture approach that offers insights into the biological process of neurite growth and inhibition,but also proposes a specific mechanism for reduced neural connectivity in the presence of amyloid-βpeptides,along with potential intervention points to restore neurite growth.Thereby,we aim to establish a culture system that has the potential to serve as an assay for measuring preclinical,predictive outcomes of drugs and their ability to promote neurite outgrowth,both generally and in a patient-specific manner.
文摘With the rise of new media and short videos shaping the new communication environment,rural culture has been mediated and transformed to spread to a wider region.As a cultural achievement of rural revitalization,“Village BA”(Village Basketball Association)demonstrates Chinese modernization.The use of short videos,mass rural movements,and“spectacle”spaces to attract spectators has become a key issue in the dissemination of rural culture.
文摘The three-dimensional(3D)cell culture system has garnered significant attention in recent years as a means of studying cell behavior and tissue development,as opposed to traditional two-dimensional cultures.These systems can induce specific cell reactions,promote specific tissue functions,and serve as valuable tools for research in tissue engineering,regenerative medicine,and drug discovery.This paper discusses current developments in the field of three-dimensional cell culture and the potential applications of 3D type 1 collagen gels to enhance the growth and maturation of dendritic cells.
文摘In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were induced to form callus when cultured on vermicompost extract media along with coelomic fluid. Suspension medium was developed using vermicompost extract and coelomic fluid in 3:1 ratio. Phytochemical analysis of the alkaloid berberine was confirmed from callus, suspension cell culture and suspension medium by Thin Layer Chromatography and High Performance Liquid Chromatography. Vermicompost and its extracts with coelomic fluid have shown maximum (100 per cent) response of callus induction. Callus mass enlarged with increasing concentration of coelomic fluid and callus growth was assessed from the biomass. Incubation of culture tubes in dark supported callus development significantly. The Rf value of 0.36 confirmed the presence of berberine by Thin Layer Chromatography. Qualitative analysis confirmed the presence of alkaloid berberine with the retention time of 2.8 minutes similar to that of standard reference sample from Sigma chemicals, USA. The suspension medium turned deep yellow because of the release of the alkaloid. Vermicompost and its extracts along with coelomic fluid have shown the economical approach for micropropagation of economically and medicinally important plants.
基金the research findings of the research project of Sichuan International Studies University(Serial number:Sisu2019016)Project Title The Impact of New Media Hotspots on Urban Tourism
文摘Nowadays,the communication of traditional culture is facing various challenges,including limitations of traditional culture itself,decrease of inheritors,poor construction of structure,and heterization of connotation,as well as conflicts between Chinese and foreign cultures in the context of globalization.New media has provided more opportunities for the communication of outstanding Chinese traditional culture.This paper analyzed the role of new media in completing the structure and expanding the scope of communication of outstanding Chinese traditional culture,and proposed strategy for optimizing the communication route from the perspective of improving environment,changing content,and adjusting strategy.The contributions of new media to the communication of Chinese traditional culture should be highly appreciated.