| 昆明小鼠胚胎干细胞滋养层制备条件的实验研究 |
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| 引用本文： | 黄林,莫曾南,覃敏.昆明小鼠胚胎干细胞滋养层制备条件的实验研究[J].现代生物医学进展,2010,10(1). |
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| 作者姓名： | 黄林 莫曾南 覃敏 |
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| 作者单位： | 广西医科大学第一附属医院泌尿外科,广西医科大学泌尿科学研究所,广西,南宁,530021 |
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| 基金项目： | 广西壮族自治区研究生科研创新项目(2009105981002M197); 广西科学研究与技术开发计划课题(桂科攻0779002,桂科攻0322025-20) |
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| 摘 要： | 目的:建立小鼠胚胎成纤维细胞(MEFS)滋养层,用于昆明小鼠胚胎干细胞的培养。方法:取妊娠13.5的胎鼠,采用组织消化法分离培养出原代成纤维细胞,对MEFs的生长形态、生长曲线及分裂指数进行观察;MTT法筛选丝裂霉素C(MMC)作用的最佳浓度和时间;取妊娠3.5d的囊胚在经MMC处理的饲养层上培养,观察胚胎干细胞集落生成情况。结果:MEFS为一种贴壁生长且增殖速度较快的细胞,第三代细胞增殖旺盛,第5代以后细胞开始变形并趋于衰老;MMC能抑制胚胎成纤维细胞的增殖,最佳的作用浓度和时间是10ug/ml作用2.5～4h,20ug/ml作用1-2.5h。妊娠3.5d小鼠囊胚在饲养层上培养能形成典型的"鸟巢"状干细胞集落,并可维持胚胎干细胞的正常形态且不发生分化。结论:这种方法制备的滋养细胞层适用于胚胎干细胞的培养。
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| 关 键 词： | 胚胎成纤维细胞 胚胎干细胞 滋养层 |
Study on Preparation of Feeder Layers for Embryonic Stem Cells Of Kunming Mouse |
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| HUANG Lin,MO Zeng-nan,QIN Min.Study on Preparation of Feeder Layers for Embryonic Stem Cells Of Kunming Mouse[J].Progress in Modern Biomedicine,2010,10(1). |
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| Authors: | HUANG Lin MO Zeng-nan QIN Min |
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| Institution: | HUANG Lin,MO Zeng-nan,QIN Min(Department of Urology,Institute of Urology , Nephrology,First Affiliated Hospital,Guang Xi Medical University,Guangxi Nanning,530021) |
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| Abstract: | Objective:To establish mouse embryonic fibroblasts as the feeder layers for supporting mouse embryonic stem cells in vitro.Methods:The mouse primary embryonic fibroblast cell(PMEFs) were isolated from the mouse embryos of 13.5 day gestation through primary tissue digestion.The morphology,growth curve and Mitotic Index of MEFs in vitro was investigated.The optional concentration of mitomycin C and the time treated with mitomycin C on MEFs were determined by MTT assay.Murine blastocysts(3.5 dpc) were cultured... |
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| Keywords: | Embryonic fibroblasts Embryonic stem cell Feeder layer |
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