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Current status and challenges for cell-cultured milk technology: a systematic review
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作者 Hyuk Cheol Kwon Hyun Su Jung +1 位作者 Vahinika Kothuri Sung Gu Han 《Journal of Animal Science and Biotechnology》 SCIE CAS CSCD 2024年第5期1778-1792,共15页
Cellular agriculture is an innovative technology for manufacturing sustainable agricultural products as an alternative to traditional agriculture.While most cellular agriculture is predominantly centered on the produc... Cellular agriculture is an innovative technology for manufacturing sustainable agricultural products as an alternative to traditional agriculture.While most cellular agriculture is predominantly centered on the production of cultured meat,there is a growing demand for an understanding of the production techniques involved in dairy products within cellular agriculture.This review focuses on the current status of cellular agriculture in the dairy sector and technical challenges for cell-cultured milk production.Cellular agriculture technology in the dairy sector has been classified into fermentation-based and animal cell culture-based cellular agriculture.Currently,various companies synthesize milk components through precision fermentation technology.Nevertheless,several startup companies are pursuing animal cell-based technology,driven by public concerns regarding genetically modified organisms in precision fermentation technology.Hence,this review offers an up-to-date exploration of animal cell-based cellular agriculture to produce milk components,specifically emphasizing the structural,functional,and productive aspects of mammary epithelial cells,providing new information for industry and academia. 展开更多
关键词 cell culture system cell-cultured milk Mammary epithelial cells Precision fermentation
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MACS-W:A modified optical clearing agent for imaging 3D cell cultures
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作者 Xiang Zhong Chao Gao +6 位作者 Hui Li Yuening He Peng Fei Zaozao Chen Zhongze Gu Dan Zhu Tingting Yu 《Journal of Innovative Optical Health Sciences》 SCIE EI CSCD 2024年第2期24-34,共11页
Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible... Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible to probe the complexity of 3D cell cultures but are limited by the inherent opaqueness.While tissue optical clearing methods have emerged as powerful tools for investigating whole-mount tissues in 3D,they often have limitations,such as being too harsh for fragile 3D cell cultures,requiring complex handling protocols,or inducing tissue deformation with shrinkage or expansion.To address this issue,we proposed a modified optical clearing method for 3D cell cultures,called MACS-W,which is simple,highly efficient,and morphology-preserving.In our evaluation of MACS-W,we found that it exhibits excellent clearing capability in just 10 min,with minimal deformation,and helps drug evaluation on tumor spheroids.In summary,MACS-W is a fast,minimally-deformative and fluorescence compatible clearing method that has the potential to be widely used in the studies of 3D cell cultures. 展开更多
关键词 Tissue optical clearing 3D cell cultures IMAGING
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3D Collagen Gels:A Promising Platform for Dendritic Cell Culture in Biomaterials Research
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作者 Kirubanandan Shanmugam 《Proceedings of Anticancer Research》 2024年第4期124-134,共11页
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
关键词 Three-dimensional cell culture Dendritic cells Type 1 collagen gels Bovine tendons and rat tails
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In Vitro Development of Tobacco Primary Endosperm Cells in Microculture 被引量:1
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作者 李师弢 房克凤 杨弘远 《Acta Botanica Sinica》 CSCD 2000年第5期542-544,共3页
用酶解_研磨法分离出烟草 (NicotianatabacumL .)受精后胚囊和初生胚乳细胞进行微室饲养培养。培养基为Km8p附加各种其他成分 ,饲养细胞为分裂旺盛的烟草叶肉原生质体 ,在 2 5℃下静止暗培养。培养 3d后 ,初生胚乳细胞开始第一次分裂 ,... 用酶解_研磨法分离出烟草 (NicotianatabacumL .)受精后胚囊和初生胚乳细胞进行微室饲养培养。培养基为Km8p附加各种其他成分 ,饲养细胞为分裂旺盛的烟草叶肉原生质体 ,在 2 5℃下静止暗培养。培养 3d后 ,初生胚乳细胞开始第一次分裂 ,继续分裂至 14d时形成大的细胞团。首次报道了双子叶植物初生胚乳细胞的离体发育。 展开更多
关键词 primary endosperm cell in vitro development icotiana tabacum
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Improvement of in vitro Cytoplasmic Maturation of Denuded Porcine Oocytes by Coculture with Follicular Mural Granulosa Cells
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作者 郇延军 解炳腾 +2 位作者 朱江 孔庆然 刘忠华 《Agricultural Science & Technology》 CAS 2012年第6期1368-1372,共5页
[Objective] This study aimed to improve the in vitro maturation quality of denuded porcine oocytes and provide scientific basis for establishing a stable and efficient denuded oocyte culture system. [Method] The first... [Objective] This study aimed to improve the in vitro maturation quality of denuded porcine oocytes and provide scientific basis for establishing a stable and efficient denuded oocyte culture system. [Method] The first polar body extrusion rate, oocyte glutathione (GSH) content, positive rate of brilliant cresyl blue (BCB) staining and development potential of activated oocytes or fertilized oocytes were employed as main indicators to investigate the effects of follicular mural granulosa cell (MGC) coculture on cytoplasmic maturation of cumulus cell-removal oocytes (Denuded Oocyte, DO). [Result] According to in vitro maturation results, compared with DO group, the first polar body extrusion rate of porcine oocytes in DO+MGC group was not significantly different, but the nuclear maturation process was improved and was more similar to that in COC (cumulus-oocyte complex) group. Detection of GSH content in mature oocytes showed that there was no significant difference between DO+ MGC group (optical density of 1 053.67) and COC group (optical density of 1 426.00) or between DO+MGC group and COC+GC group (optical density of 1 541.00), however, GSH content in mature oocytes of DO group (optical density of 724.67) was significantly lower than that of COC group and COC+GC group (P0.05). Detection of glucose-6-phosphate dehydrogenase (G6PDH) activity showed that there was no significant difference in BCB positive oocyte rate between DO +MGC group (88.26% ) and COC group (92.75%) or between DO+MGC group and DO group (82.86% ), however, BCB positive oocyte rate of DO group was significantly lower than that of COC group (P0.05). Furthermore, the cleavage rate and blastocyst rate of activated mature oocytes derived from DO +MGC group (94.98% and 43.67% , respectively) were significantly higher than those from DO group (52.54% and 8.97%, respectively) (P0.05), and were not significantly different compared with those from COC group (97.11% and 38.30%, respectively). In addition, the cleavage rate of fertilized oocytes derived from DO+MGC group (72.65%) showed no significant difference compared with that from DO group (63.59%), but the blastocyst rate of DO+MGC group was significantly higher than that of DO group (9.88%) (P0.05). [Conclusion] MGC coculture can significantly improve the in vitro cytoplasmic maturation quality of denuded porcine oocytes, thereby enhancing the subsequent developmental potential. 展开更多
关键词 Denuded oocytes Mural granulosa cells in vitro maturation Cytoplasmic maturation PORCINE
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Long-term in-vitro culture and subculture of the hemocytes of swimming crab Portunus trituberculatus
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作者 Liwen GUO Yaqi ZHAO Huarong GUO 《Journal of Oceanology and Limnology》 SCIE CAS CSCD 2023年第5期1918-1939,共22页
Crab cell line,especially continuous crab cell line,can provide us a useful tool for studies on the virology,immunology,and molecular biology of crabs.However,no continuous crab cell line has been available due to the... Crab cell line,especially continuous crab cell line,can provide us a useful tool for studies on the virology,immunology,and molecular biology of crabs.However,no continuous crab cell line has been available due to the lacking of suitable medium and the occurrence of mitosis-arrest.In this study,long-term in vitro culture conditions for both two-(2D)and three-dimensions(3D)were successfully developed for the circulating hemocytes of swimming crab Portunus trituberculatus,designated as PTH cells.In 2D culture,a novel crab basic medium in osmolarity of 990–1100 mOsm/kg was optimized for the first time,which is different from Leibovitz's L-15 medium in mainly the components of amino acids,containing double strengths of the contents of free amino acid mixture in the crab serum.Then an optimal crab growth medium was developed by supplementing 5%fetal bovine serum,50-g/L yeast extract powder,20-μg/L basic fibroblast growth factor and epidermal growth factor into the optimal crab basic medium,and found that it could support a long-term survival of PTH cells in a healthy monolayer up to 347 days and partially break through the mitosis-arrest of crab cells evidenced by the obvious increase of proliferating potential detected in the 10-d primarily cultured PTH cells.These 2D cultured PTH cells could be successfully sub-cultured for 11 times by physical flushing method and well cryopreserved in liquid nitrogen.In 3D culture,using the same crab growth medium,the PTH cell aggregates could be easily formed and healthily maintained on the surface of solidified Matrigel or in the ultra-low-attachment plate with a survival rate of 50%–60%on Day 103.This work largely improved the primary culture and subculture of crab cells and will facilitate the establishment of continuous crab cell line. 展开更多
关键词 CRAB Portunus trituberculatus HEMOCYTE long-term cell culture SUBculture 3D culture
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Culture and identification of neonatal rat brain-derived neural stem cells
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作者 Qing-Zhong Zhou Xiao-Lan Feng +4 位作者 Xu-Feng Jia Nurul Huda Binti Mohd Nor Mohd Hezery Bin Harun Da-Xiong Feng Wan Aliaa Wan Sulaiman 《World Journal of Stem Cells》 SCIE 2023年第6期607-615,共9页
BACKGROUND Timing of passaging,passage number,passaging approaches and methods for cell identification are critical factors influencing the quality of neural stem cells(NSCs)culture.How to effectively culture and iden... BACKGROUND Timing of passaging,passage number,passaging approaches and methods for cell identification are critical factors influencing the quality of neural stem cells(NSCs)culture.How to effectively culture and identify NSCs is a continuous interest in NSCs study while these factors are comprehensively considered.AIM To establish a simplified and efficient method for culture and identification of neonatal rat brain-derived NSCs.METHODS First,curved tip operating scissors were used to dissect brain tissues from new born rats(2 to 3 d)and the brain tissues were cut into approximately 1 mm^(3)sections.Filter the single cell suspension through a nylon mesh(200-mesh)and culture the sections in suspensions.Passaging was conducted with TrypLTM Express combined with mechanical tapping and pipetting techniques.Second,identify the 5th generation of passaged NSCs as well as the revived NSCs from cryopreservation.BrdU incorporation method was used to detect self-renew and proliferation capabilities of cells.Different NSCs specific antibodies(anti-nestin,NF200,NSE and GFAP antibodies)were used to identify NSCs specific surface markers and muti-differentiation capabilities by immunofluorescence staining.RESULTS Brain derived cells from newborn rats(2 to 3 d)proliferate and aggregate into spherical-shaped clusters with sustained continuous and stable passaging.When BrdU was incorporated into the 5th generation of passaged cells,positive BrdU cells and nestin cells were observed by immunofluorescence staining.After induction of dissociation using 5%fetal bovine serum,positive NF200,NSE and GFAP cells were observed by immunofluorescence staining.CONCLUSION This is a simplified and efficient method for neonatal rat brain-derived neural stem cell culture and identification. 展开更多
关键词 Neonatal rats Brain-derived neural stem cells culture IDENTIFICATION
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An Innovative Design of Incubator Structure for Cell Culture
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作者 Shanshan HE Zhongwei CHEN +2 位作者 Ruonan HE Shiyi WU Qihuang LIN 《Medicinal Plant》 CAS 2023年第3期105-107,共3页
In view of the problems of the traditional cell incubator,such as the small range of cell culture types,the inability to adjust the internal space of the incubator according to needs,and the inconvenient sampling,this... In view of the problems of the traditional cell incubator,such as the small range of cell culture types,the inability to adjust the internal space of the incubator according to needs,and the inconvenient sampling,this study innovatively designed a cell incubator structure.It proposed a new design concept that can solve the above-mentioned shortcomings.The cell incubator after the new structural modification can adjust the internal space structure of cell culture by setting the bolt-fixed connection between the fixed plate and the vessel divider.It realizes the cultivation of various cells through refrigeration modules and heating modules.Through setting a sampling hole in the glass inner door,it is favorable for operators to take samples,making cell culture more convenient and efficient. 展开更多
关键词 cell incubator Innovative design cell culture
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Repetitive administration of cultured human CD34+cells improve adenine-induced kidney injury in mice
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作者 Takayasu Ohtake Shoichi Itaba +9 位作者 Amankeldi A Salybekov Yin Sheng Tsutomu Sato Mitsuru Yanai Makoto Imagawa Shigeo Fujii Hiroki Kumagai Masamitsu Harata Takayuki Asahara Shuzo Kobayashi 《World Journal of Stem Cells》 SCIE 2023年第4期268-280,共13页
BACKGROUND There is no established treatment to impede the progression or restore kidney function in human chronic kidney disease(CKD).AIM To examine the efficacy of cultured human CD34+cells with enhanced proliferati... BACKGROUND There is no established treatment to impede the progression or restore kidney function in human chronic kidney disease(CKD).AIM To examine the efficacy of cultured human CD34+cells with enhanced proliferating potential in kidney injury in mice.METHODS Human umbilical cord blood(UCB)-derived CD34+cells were incubated for one week in vasculogenic conditioning medium.Vasculogenic culture significantly increased the number of CD34+cells and their ability to form endothelial progenitor cell colony-forming units.Adenineinduced tubulointerstitial injury of the kidney was induced in immunodeficient non-obese diabetic/severe combined immunodeficiency mice,and cultured human UCB-CD34+cells were administered at a dose of 1×106/mouse on days 7,14,and 21 after the start of adenine diet.RESULTS Repetitive administration of cultured UCB-CD34+cells significantly improved the time-course of kidney dysfunction in the cell therapy group compared with that in the control group.Both interstitial fibrosis and tubular damage were significantly reduced in the cell therapy group compared with those in the control group(P<0.01).Microvasculature integrity was significantly preserved(P<0.01)and macrophage infiltration into kidney tissue was dramatically decreased in the cell therapy group compared with those in the control group(P<0.001).CONCLUSION Early intervention using human cultured CD34+cells significantly improved the progression of tubulointerstitial kidney injury.Repetitive administration of cultured human UCB-CD34+cells significantly improved tubulointerstitial damage in adenine-induced kidney injury in mice via vasculoprotective and anti-inflammatory effects. 展开更多
关键词 Chronic kidney disease CD34+cell ADENINE Tubulointerstitial injury Quality and quantity control culture Umbilical cord blood
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Optimization of Culture Medium and Transfection Method for Head and Neck Squamous Cell Carcinoma Organoids
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作者 Zhongheng HUANG Xi YAO +2 位作者 Qi LIU Ying XIE Zhengbo WEI 《Medicinal Plant》 CAS 2023年第3期100-104,共5页
[Objectives] To optimize the culture medium for head and neck squamous cell carcinoma patient-derived organoid and screen suitable cytokines;compare the transfection efficiency of direct transfection and short-term su... [Objectives] To optimize the culture medium for head and neck squamous cell carcinoma patient-derived organoid and screen suitable cytokines;compare the transfection efficiency of direct transfection and short-term suspension transfection for organoid in matrigel. [Methods] Advanced DMEM/F12 medium, GlutaMax and HEPES buffer, nicotinamide, N-acetylcysteine, B27, A83-01, EGF, Y-27632 and Primocin primary cell antibiotics were prepared. On this basis, fibroblast growth factor 10(FGF10), Neuregulin 1, Noggin and R-spondin-1 were added in turn to prepare the selection medium, and the organoid diameter was used as the evaluation index to evaluate the effect of organoid medium. Using lentivirus, mCherry red fluorescent protein was transfected into HNSCC—PDO in different ways, and the transfection effect was evaluated by the fluorescence intensity of organoid sphere. [Results] Nrg1 Noggin and R-Spondin-1 promoted the growth of head and neck squamous cell carcinoma sphere(P<0.05) while FGF10 did not significantly promote the growth of head and neck squamous cell carcinoma sphere(P>0.05). Compared with direct transfection, short-term suspension transfection had higher transfection efficiency for HNSCC—PDO in matrigel. [Conclusions] R-Spondin-1 Nrg1 and Noggin may be the key cytokines in culture of HNSCC—PDO whereas FGF10 played an insignificant role in this study. Short-term suspension transfection could improve the transfection efficiency of lentivirus to HNSCC—PDO. 展开更多
关键词 Head and neck squamous cell carcinoma Organoid culture Organoid transfection
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Sequential extraction of RNA,DNA and protein from cultured cells of the same group
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作者 Ying-Yu Cui 《World Journal of Methodology》 2023年第5期484-491,共8页
BACKGROUND Efficient extraction of nucleic acids and proteins(ENAP)from cells is a prerequisite for precise annotation of gene function,and has become laboratory routine for revealing the mysteries of life.However,cel... BACKGROUND Efficient extraction of nucleic acids and proteins(ENAP)from cells is a prerequisite for precise annotation of gene function,and has become laboratory routine for revealing the mysteries of life.However,cell samples are often from different culture dishes,resulting in inevitable experimental errors and sometimes poor repeatability.AIM To explore a method to improve the efficiency of ENAP,minimizing errors in ENAP processes,enhancing the reliability and repeatability of subsequent experimental results.METHODS A protocol for the sequential isolation of RNA,DNA,and proteins from the same cultured HepG2 cells using RNAzol reagent is presented here.The first step involves culturing HepG2 cells to the exponential phase,followed by the sequential isolation of RNA,DNA,and proteins from the same cultured cells in the second step.The yield of nucleic acids and proteins is detected in the third step,and their purity and integrity are verified in the last step.RESULTS The procedure takes as few as 3-4 d from the start to quality verification and is highly efficient.In contrast to the existing kits and reagents,which are primarily based on independent isolation,this RNAzol reagent-based method is characterized by the sequential isolation of RNA,DNA,and proteins from the same cells,and therefore saves time,and has low cost and high efficiency.CONCLUSION The RNA,DNA,and proteins isolated using this method can be used for reverse transcription-polymerase chain reaction,polymerase chain reaction,and western blotting,respectively. 展开更多
关键词 Sequential extraction Ribonucleic acid Deoxyribonucleic acid PROTEIN cultured cells
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In vitro culture and differentiation of rat embryonic midbrain-derived neural stem cells 被引量:19
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作者 Xingli Deng Ruen Liu +5 位作者 Zhongtang Feng Jing Guo Wu Wang Deqiang Lei Hongyan Li Zhihua Chen 《Neural Regeneration Research》 SCIE CAS CSCD 2008年第11期1241-1244,共4页
BACKGROUND: Midbrain-derived neural stem cells (mNSCs) can differentiate into functional mature dopaminergic neurons. The mNSCs are considered the ideal choice for cell therapy of Parkinson's disease. OBJECTIVE: ... BACKGROUND: Midbrain-derived neural stem cells (mNSCs) can differentiate into functional mature dopaminergic neurons. The mNSCs are considered the ideal choice for cell therapy of Parkinson's disease. OBJECTIVE: To isolate rat embryonic mNSCs and to observe the differentiation characteristics of mNSCs induced by cell growth-promoting factors. DESIGN, TIME AND SETTING: An in vitro cell culture study based on the molecular biology of nerve cells was carried out at the Institute of Clinical Medicine, China-Japan Friendship Hospital (China) from March to November 2007. MATERIALS: Sprague Dawley rats at embryonic day 14 were used in this study. Nestin antibody, β-Ⅲ tubulin antibody, glial fibrillary acidic protein (GFAP) antibody and cyclic nucleotide 3'-phosphohydrolase (CNPase) antibody were provided by Abcam; DMEM/F12 medium and N2 supplement were provided by Invitrogen; epidermal growth factor (EGF) and fibroblast growth factor-2 (FGF2) were provided by R&D Systems. METHODS: The ventral mesencephalon was dissected from embryonic day 14 rat embryos. By trypsin digestion and mechanical separation, the brain tissue was triturated into a fine single-cell suspension. The cells were cultured in 5 mL serum-free medium containing DMEM/FI 2, 1% N: supplement, 20 ng/mL EGF and FGF2. The mNSCs at the third generation were coated with 10ug/mL polylysine and induced to differentiate in the DMEM/F12 supplemented with 1% fetal bovine serum and 1% N2. MAIN OUTCOME MEASURES: The neural spheres of the third passage were identified by nestin immunofluorescence; at the same time, the cells were induced to differentiate, and the types of differentiated cell were identified by immunofluorescence for β Ⅲ tubulin, GFAP and CNPase. RESULTS: Seven days after primary culture, a great many neurospheres could be obtained by successive pasage. Immunofluorescence assays showed that the neurospheres were nestin positive, and after differentiation, the cells expressed GFAP, CNPase and β -Ⅲ-tubulin. CONCLUSION: Embryonic day 14 rat mNSCs can differentiate into neuron-like cells and glial cells following induction by EGF, FGF2 and N: additive. 展开更多
关键词 neural stem cells cell differentiation in vitro rat embryonic midbrain
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Three-dimensional cell culture systems as an in vitro platform for cancer and stem cell modeling 被引量:10
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作者 Nipha Chaicharoenaudomrung Phongsakorn Kunhorm Parinya Noisa 《World Journal of Stem Cells》 SCIE 2019年第12期1065-1083,共19页
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
关键词 Three-dimensional cultureS CANCER Stem cells Disease MODELING In vitro screening PLATFORM
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In vitro Culture of Bone Marrow Mesenchymal Stem Cells in Rats and Differentiation into Retinal Neural-like Cells 被引量:6
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作者 孙旭芳 姜焕荣 杨红 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2007年第5期598-600,共3页
In order to study the in vitro culture and expansion of bone marrow mesenchymal stem cells in rats (rMSCs) and the possibility of rMSCs differentiation into retinal neural cells, the bone marrow-derived cells in SD ... In order to study the in vitro culture and expansion of bone marrow mesenchymal stem cells in rats (rMSCs) and the possibility of rMSCs differentiation into retinal neural cells, the bone marrow-derived cells in SD rats were isolated and cultured in vitro. The retinal neural cells in SD rats were cultured and the supernatants were collected to prepare conditioned medium. The cultured rMSCs were induced to differentiate by two steps. Immunofluorescence method and anti-nestin, anti-NeuN, anti-GFAP and anti-Thyl. 1 antibodies were used to identify the cells derived from the rMSCs. The results showed that the in vitro cultured rMSCs grew well and expanded quickly. After induction with two conditioned media, rMSCs was induced to differentiate into neural progenitor cells, then into retinal neural-like cells which were positive for nestin, NeuN, GFAP and Thyl. 1 detected by fluorescence method. The findings suggested that rMSCs could be culture and expanded in vitro, and induced to differentiate into retinal neural-like cells. 展开更多
关键词 bone marrow stem cells cell culture DIFFERENTIATION retinal neural cell
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Progress in in vitro culture and gene editing of porcine spermatogonial stem cells 被引量:3
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作者 Yi-Zhuo Sun Si-Tong Liu +1 位作者 Xiao-Meng Li Kang Zou 《Zoological Research》 SCIE CAS CSCD 2019年第5期343-348,共6页
Research on in vitro culture and gene editing of domestic spermatogonial stem cells (SSCs) is of considerable interest but remains a challenging issue in animal science. In recent years, some progress on the isolation... Research on in vitro culture and gene editing of domestic spermatogonial stem cells (SSCs) is of considerable interest but remains a challenging issue in animal science. In recent years, some progress on the isolation, purification, and genetic manipulation of porcine SSCs has been reported. Here, we summarize the characteristics of porcine SSCs as well current advances in their in vitro culture, potential usage, and genetic manipulation. Furthermore, we discuss the current application of gene editing in pig cloning technology. Collectively, this commentary aims to summarize the progress made and obstacles encountered in porcine SSC research to better serve animal husbandry, improve livestock fecundity, and enhance potential clinical use. 展开更多
关键词 PROGRESS vitro culture STEM cells
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Islet separation and islet cell culture in vitro from human embryo-pancreas 被引量:3
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作者 Yuan Z Wu GY +2 位作者 He YS Shao CM Zhan Y 《World Journal of Gastroenterology》 SCIE CAS CSCD 1999年第5期458-460,共3页
关键词 cell culture ISLET cell ISLETS of LANGERHANS transplantetion pancreas/ EMBRYOLOGY
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In Vitro Invasive Pattern of Hepatocellular Carcinoma Cell Line HCCLM9 Based on Three-dimensional Cell Culture and Quantum Dots Molecular Imaging 被引量:7
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作者 方敏 彭春伟 +2 位作者 刘少平 袁静萍 李雁 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2013年第4期520-524,共5页
Summary: This study aimed to establish a new in vitro three-dimensional (3D) cell culture and use quantum dots (QDs) molecular imaging to examine the invasive behaviors of hepatocellular carcinoma (HCC) cells. ... Summary: This study aimed to establish a new in vitro three-dimensional (3D) cell culture and use quantum dots (QDs) molecular imaging to examine the invasive behaviors of hepatocellular carcinoma (HCC) cells. Each well of the 24-well cell culture plate was cover-slipped. Matrigel diluted with se- rum-free DMEM was added and HCCLM9 cells were cultured on the Matrigel. The cell morphological and cell growth characteristics were observed by inverted microscopy and laser confocal microscopy at different culture time. Cell invasive features were monitored by QDs-based real-time molecular imaging techniques. The results showed that on this 3D cell culture platform, HCCLM9 cells exhibited typical multi-step invasive behaviors, including reversion of cell senescence, active focal proliferation and dominant clones invasion. During the process, cells under 3D cell culture showed biological behaviors of spatio-temporal characteristics. Cells first merged on the surface of matrix, then gradually infiltrated and migrated into deep part of matrix, presenting polygonal morphology with stretched protrusions, forming tubular, annular and even network structure, which suggested that HCC cells have the morpho- logical basis for vasculogenic mimicry. In addition, small cell clones with their edges well-circumscribed in early stage, progressed into a large irregular clone with ill-defined edge, while the other cells developed invadopodia. And QDs probing showed MT1-MMP was strongly expressed in the invadopodia. These findings indicate that a novel 3D cell culture platform has been successfully estab- lished, which can mimic the in vivo tumor microenvironment, and when combined with QDs-based mo- lecular imaging, it can help to better investigate the invasive behaviors of HCC cells. 展开更多
关键词 3D cell culture tumor microenvironment tumor invasion quantum dots
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Biological Characteristics of Human Bone Marrow Mesenchymal Stem Cell Cultured in Vitro 被引量:5
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作者 法宪恩 王利霞 +3 位作者 侯剑峰 张瑞成 王海永 杨辰垣 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2005年第3期307-309,共3页
Some biological characteristics of human bone marrow mesenchymal stem cells (MSCs) cultured in vitro were observed. hMSCs were isolated from bone marrow and purified by density gradient centrifugation method, and then... Some biological characteristics of human bone marrow mesenchymal stem cells (MSCs) cultured in vitro were observed. hMSCs were isolated from bone marrow and purified by density gradient centrifugation method, and then cultured in vitro. The proliferation and growth characteristics of hMSCs were observed in primary and passage culture. MSCs of passage 3 were examined for the purify by positive rate of CD29 and CD44 through flow cytometry. Human bone marrow MSCs showed active proliferation capacity in vitro. The purify of MSCs separated by our method was higher than 90 %. It was concluded that hMSCs have been successfully cultured and expanded effectively. It provided a foundation for further investigation and application of MSCs. 展开更多
关键词 mesenchymal stem cell bone marrow cell culture
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Dissociation, culture and morphologic changes of interstitial cells of Cajal in vitro 被引量:10
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作者 Chun-XueLi Bao-HuaLiu Wei-DongTong Lian-YangZhang Yan-PingJiang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第18期2838-2840,共3页
AIM: To study the method of dissociation, culture and investigate its morphologic changes in vitro of interstitial cells of Cajal (ICC).METHODS: Enzymatic digestion and Ficoll density centrifugation were used to disso... AIM: To study the method of dissociation, culture and investigate its morphologic changes in vitro of interstitial cells of Cajal (ICC).METHODS: Enzymatic digestion and Ficoll density centrifugation were used to dissociate ICC from the ileal segment of mice. Factors including contamination, Ca2+, Mg2+ and collagenase, and stem cell factor, etc., were investigated.ACK2, the antibody of c-kit, was used to identify the cultured ICC. Both light microscope and fluorescence microscope were used to observe the changes of ICC in vitro.RESULTS: The method for dissociation and culture of ICC in vitro was successfully established. After 24 h, cultured ICC exhibited a few axis-cylinders, and longer axis-cylinders were observed to form synapse of each other after 3 d. More widespread connections formed within 7 d in vitro. The changes of its morphologic character were obvious within 7 d; however, there were no obvious morphologic changes after 30 d.CONCLUSION: Many factors can influence the dissociation and culture of ICC. 展开更多
关键词 Interstitial cells of Cajal culture Morphologic change
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Isolation of Cultured Endothelial Progenitor Cells in vitro from PBMCs and CD133^+ Enriched Cells 被引量:2
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作者 郑伟红 万亚峰 +4 位作者 马小鹏 李兴睿 杨志芳 殷茜 易继林 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2010年第1期18-24,共7页
Two isolation methods for sorting of endothelial progenitor cells(EPCs):from peripheral blood mononuclear cells(PBMCs)and CD133+ enriched cells were compared,by defining the cell morphology,phenotype,reproductive acti... Two isolation methods for sorting of endothelial progenitor cells(EPCs):from peripheral blood mononuclear cells(PBMCs)and CD133+ enriched cells were compared,by defining the cell morphology,phenotype,reproductive activities and function in vitro,to provide a reference for clinical application of EPCs.PBMCs from healthy subjects were used either directly for cell culture or for CD133+ sorting.The two groups of cells were cultured in complete medium 199(M199)for 7 to 14 days and the phenotypes of EPCs were an... 展开更多
关键词 endothelial progenitor cells cell culture MACS
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