“The Belt and Road”cooperation initiative contains Chinese excellent traditional cultural ideas,and is deeply rooted in Chinese excellent traditional culture.The proposal of“the Belt and Road”cooperation initiativ...“The Belt and Road”cooperation initiative contains Chinese excellent traditional cultural ideas,and is deeply rooted in Chinese excellent traditional culture.The proposal of“the Belt and Road”cooperation initiative is not only the strategic need to implement the“going out”of Chinese culture,but also the need to promote cultural exchanges and mutual learning among countries along“the Belt and Road”,and more importantly,the practical consideration of building a community of shared future for mankind with cultural identity.Confucius classroom,as one of the important platforms to realize the“going out”of Chinese culture,shows the“image of China”and enhances the“soft power”of Chinese culture,integrates and configures Chinese teaching and Chinese cultural resources through cooperation,and plays an important role in telling Chinese stories,spreading Chinese voice,enhancing cultural mutual trust and people-to-people contacts as well as the awareness of a community of shared future for mankind.Taking the Confucius classroom jointly built by Huanggang Normal University in China and Sabaragamuwa University in Sri Lanka as an example,this paper explores some effective transmission ways for spreading Chinese excellent traditional culture in Sri Lankan Confucius classroom under the background of“the Belt and Road”cooperation initiative,aiming to give full play to the role of Confucius classroom in cultural exchanges and enhance inter-school exchanges and cooperation between China and Sri Lanka in terms of cultural mutual trust and respect.展开更多
We show that giant asymmetric transmission and optical rotation for linear polarizations can be achieved by a chiral three-dimensional metamaterial composed of L-shaped and C-shaped metallic particles. Numerical calcu...We show that giant asymmetric transmission and optical rotation for linear polarizations can be achieved by a chiral three-dimensional metamaterial composed of L-shaped and C-shaped metallic particles. Numerical calcu- lations on the electric field distributions indicate that the coupling between the electric dipolar and quadrupolar resonances in the L- and C-shaped metallic particles contributes to these effects.展开更多
The 5,000-year-history of Chinese culture has produced very rich cultural traditions.However,with the deepening of reform and opening up,some foreign cultures and theories have had a significant impact on the Chinese ...The 5,000-year-history of Chinese culture has produced very rich cultural traditions.However,with the deepening of reform and opening up,some foreign cultures and theories have had a significant impact on the Chinese traditional culture,which has severely restricted its further development.Moreover,there are still many deficiencies in the process of China’s cultural transmission,which leads to the stagnant efficiency and level of cultural communication in the intercultural communication environment.From the perspective of intercultural communication,based on the analysis of the problems existing in China’s cultural transmission,this paper puts forward some suggestions and strategies to promote China’s cultural transmission,and facilitate the development of Chinese culture in the context of the interweaving of world culture.展开更多
In this work,the three-dimensional(3 D)propagation behaviors in the nonlinear phononic crystal and elastic wave metamaterial with initial stresses are investigated.The analytical solutions of the fundamental wave and ...In this work,the three-dimensional(3 D)propagation behaviors in the nonlinear phononic crystal and elastic wave metamaterial with initial stresses are investigated.The analytical solutions of the fundamental wave and second harmonic with the quasilongitudinal(qP)and quasi-shear(qS_(1) and qS_(2))modes are derived.Based on the transfer and stiffness matrices,band gaps with initial stresses are obtained by the Bloch theorem.The transmission coefficients are calculated to support the band gap property,and the tunability of the nonreciprocal transmission by the initial stress is discussed.This work is expected to provide a way to tune the nonreciprocal transmission with vector characteristics.展开更多
This article,based on the English version of Li Bai's Classical Chinese Poem“Bring in the Wine”,explores the important position of cultural transmission in the English translation of the Classical Chinese Poem t...This article,based on the English version of Li Bai's Classical Chinese Poem“Bring in the Wine”,explores the important position of cultural transmission in the English translation of the Classical Chinese Poem through the comparison of the cultural translation,and finally draws the rationality and feasibility of the creative translation techniques in the English translation of the Classical Chinese Poems.It points out that the alienation of the Chinese culture's external communication has ignored the reader's cultural acceptance ability and position of the western countries and regions dominated by the foreign countries.It is believed that in a long period of time,Chinese culture needs the translation of Chinese translators to dissimilation,the translation of foreign sinologists,domestication,and alienation,to meet the needs of different types of readers and to promote the Chinese culture to go out.展开更多
International transmission of Chinese medical culture is of great importance for the worldwide spread of traditional Chinese culture, which faces many problems, like the serious impact from western medicine and other ...International transmission of Chinese medical culture is of great importance for the worldwide spread of traditional Chinese culture, which faces many problems, like the serious impact from western medicine and other traditional medicines, low professional quality of its practitioners, language barriers, etc. It's urgent to propose some practical solutions.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
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
The Belt and Road initiative has promoted the connotative development and export of Chinese culture.To ensure a good international display and going globally of Chinese culture,we could follow such paths as excavating...The Belt and Road initiative has promoted the connotative development and export of Chinese culture.To ensure a good international display and going globally of Chinese culture,we could follow such paths as excavating the rich elements of traditional Chinese culture,shaping the fine works,creating culture brands with Chinese characteristics,developing new culture brands,opening up the cultural market,strengthening cultural dialogue,innovating cultural transmission paths by gathering the cultural forces at home and abroad,and enhancing the appeal of national culture so as to increase the closeness and recognition of people along the route.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
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
The in vitro expansion of stem cells is important for their application in different life science fields such as cellular tissue and organ repair.An objective of this paper was to achieve static cell culture in vitro ...The in vitro expansion of stem cells is important for their application in different life science fields such as cellular tissue and organ repair.An objective of this paper was to achieve static cell culture in vitro through peptide hydrogel-supported microspheres(MSs).The peptides,with their gel-forming properties,microstructures,and mechanical strengths characterized,were found to have good support for the MSs and to be injectable.The internal structures of poly(L-lactic acid)microspheres(PLLA-MSs)and polystyrene microspheres(PS-MSs)made in thelaboratory were observed and statistically analyzed in terms of particle size and pore size,following which the co-cultured MSs with cells were found to have good cell adhesion.In addition,three-dimensional(3D)culturing of cells was performed on the peptide and microcarrier composite scaffolds to measure cell viability and cell proliferation.The results showed that the peptides could be stimulated by the culture medium to self-assembly form a 3D fiber network structure.Under the peptide-Ms composite scaffold-based cell culture system,further enhancement of the cell culture effect was measured.The peptide-Ms composite scaffolds have great potential for the application in 3D cell culture and in vitro cellexpansion.展开更多
基金2021 China-Sri Lanka Cultural Exchange and Economic Development Research Center Project of Huanggang Normal University:“Research on the Dissemination and Translation of Chinese Classical Poetry in Sri Lanka From the Perspective of‘the Belt and Road’”.The research results of the project were funded by the China-Sri Lanka Cultural Exchange and Economic Development Research Center of Huanggang Normal University(Fund Project No.:202126504).
文摘“The Belt and Road”cooperation initiative contains Chinese excellent traditional cultural ideas,and is deeply rooted in Chinese excellent traditional culture.The proposal of“the Belt and Road”cooperation initiative is not only the strategic need to implement the“going out”of Chinese culture,but also the need to promote cultural exchanges and mutual learning among countries along“the Belt and Road”,and more importantly,the practical consideration of building a community of shared future for mankind with cultural identity.Confucius classroom,as one of the important platforms to realize the“going out”of Chinese culture,shows the“image of China”and enhances the“soft power”of Chinese culture,integrates and configures Chinese teaching and Chinese cultural resources through cooperation,and plays an important role in telling Chinese stories,spreading Chinese voice,enhancing cultural mutual trust and people-to-people contacts as well as the awareness of a community of shared future for mankind.Taking the Confucius classroom jointly built by Huanggang Normal University in China and Sabaragamuwa University in Sri Lanka as an example,this paper explores some effective transmission ways for spreading Chinese excellent traditional culture in Sri Lankan Confucius classroom under the background of“the Belt and Road”cooperation initiative,aiming to give full play to the role of Confucius classroom in cultural exchanges and enhance inter-school exchanges and cooperation between China and Sri Lanka in terms of cultural mutual trust and respect.
基金Supported by the International Science and Technology Cooperation Program of Anhui Province under Grant No 1403062027the Anhui Provincial Natural Science Foundation under Grant Nos KJ2014A195 and 1308085MA13the Undergraduate Scientific and Technological Innovation Project under Grant Nos AH201310371022,2013jyxml36,2013ZYJS04 and FS201310371082
文摘We show that giant asymmetric transmission and optical rotation for linear polarizations can be achieved by a chiral three-dimensional metamaterial composed of L-shaped and C-shaped metallic particles. Numerical calcu- lations on the electric field distributions indicate that the coupling between the electric dipolar and quadrupolar resonances in the L- and C-shaped metallic particles contributes to these effects.
文摘The 5,000-year-history of Chinese culture has produced very rich cultural traditions.However,with the deepening of reform and opening up,some foreign cultures and theories have had a significant impact on the Chinese traditional culture,which has severely restricted its further development.Moreover,there are still many deficiencies in the process of China’s cultural transmission,which leads to the stagnant efficiency and level of cultural communication in the intercultural communication environment.From the perspective of intercultural communication,based on the analysis of the problems existing in China’s cultural transmission,this paper puts forward some suggestions and strategies to promote China’s cultural transmission,and facilitate the development of Chinese culture in the context of the interweaving of world culture.
基金Project supported by the National Natural Science Foundation of China(Nos.11922209,11991031 and 12021002)。
文摘In this work,the three-dimensional(3 D)propagation behaviors in the nonlinear phononic crystal and elastic wave metamaterial with initial stresses are investigated.The analytical solutions of the fundamental wave and second harmonic with the quasilongitudinal(qP)and quasi-shear(qS_(1) and qS_(2))modes are derived.Based on the transfer and stiffness matrices,band gaps with initial stresses are obtained by the Bloch theorem.The transmission coefficients are calculated to support the band gap property,and the tunability of the nonreciprocal transmission by the initial stress is discussed.This work is expected to provide a way to tune the nonreciprocal transmission with vector characteristics.
文摘This article,based on the English version of Li Bai's Classical Chinese Poem“Bring in the Wine”,explores the important position of cultural transmission in the English translation of the Classical Chinese Poem through the comparison of the cultural translation,and finally draws the rationality and feasibility of the creative translation techniques in the English translation of the Classical Chinese Poems.It points out that the alienation of the Chinese culture's external communication has ignored the reader's cultural acceptance ability and position of the western countries and regions dominated by the foreign countries.It is believed that in a long period of time,Chinese culture needs the translation of Chinese translators to dissimilation,the translation of foreign sinologists,domestication,and alienation,to meet the needs of different types of readers and to promote the Chinese culture to go out.
基金sponsored by academic programs of Hubei provincial department of education(15Q112)
文摘International transmission of Chinese medical culture is of great importance for the worldwide spread of traditional Chinese culture, which faces many problems, like the serious impact from western medicine and other traditional medicines, low professional quality of its practitioners, language barriers, etc. It's urgent to propose some practical solutions.
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one of the results of the project supported by National Education Science Planning Project(A study on the cross-cultural sensitivity of Xinjiang minority preparatory students—a case study of a university,No.EMA180452)
文摘The Belt and Road initiative has promoted the connotative development and export of Chinese culture.To ensure a good international display and going globally of Chinese culture,we could follow such paths as excavating the rich elements of traditional Chinese culture,shaping the fine works,creating culture brands with Chinese characteristics,developing new culture brands,opening up the cultural market,strengthening cultural dialogue,innovating cultural transmission paths by gathering the cultural forces at home and abroad,and enhancing the appeal of national culture so as to increase the closeness and recognition of people along the route.
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.
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.
基金supported by the National Key Research andDevelopment Program of China(Grant No.2021YFC2101400).
文摘The in vitro expansion of stem cells is important for their application in different life science fields such as cellular tissue and organ repair.An objective of this paper was to achieve static cell culture in vitro through peptide hydrogel-supported microspheres(MSs).The peptides,with their gel-forming properties,microstructures,and mechanical strengths characterized,were found to have good support for the MSs and to be injectable.The internal structures of poly(L-lactic acid)microspheres(PLLA-MSs)and polystyrene microspheres(PS-MSs)made in thelaboratory were observed and statistically analyzed in terms of particle size and pore size,following which the co-cultured MSs with cells were found to have good cell adhesion.In addition,three-dimensional(3D)culturing of cells was performed on the peptide and microcarrier composite scaffolds to measure cell viability and cell proliferation.The results showed that the peptides could be stimulated by the culture medium to self-assembly form a 3D fiber network structure.Under the peptide-Ms composite scaffold-based cell culture system,further enhancement of the cell culture effect was measured.The peptide-Ms composite scaffolds have great potential for the application in 3D cell culture and in vitro cellexpansion.