胎盘间充质干细胞传代后的增殖能力

发布时间：2017-12-30 17:44

本文关键词：胎盘间充质干细胞传代后的增殖能力　出处：《中国组织工程研究》2014年10期 　论文类型：期刊论文

【摘要】：背景:胎盘间充质干细胞的增殖能力比骨髓间充质干细胞增殖能力强,但是否每个代次胎盘间充质干细胞的增殖能力都相同尚无定论。目的:分析不同代次胎盘间充质干细胞的增殖能力。方法:采用酶消化法分离人胎盘间充质干细胞,并利用流式细胞仪检测干细胞表面标志物,并进行成脂成骨诱导分化。采用细胞计数和BrdU方法检测细胞的增殖能力,根据倍增时间公式计算倍增时间,并进行长期传代。结果与结论:胎盘间充质干细胞阳性表达CD44、CD90和CD105,阴性表达CD14、CD34和CD45;成脂诱导21 d后油红O染色镜下可见红色脂肪滴,成骨诱导35 d茜素红染色镜下可见钙盐结节。细胞从第1代开始计数,直到第15代共获得3.5×1011个细胞。P5代比P10和P15代胎盘间充质干细胞的增殖能力强,倍增时间短。长期传代,胎盘间充质干细胞传代到第25代时,其中3例标本细胞形态完全发生改变,呈铺路石样;另外2例标本几乎无贴壁细胞。提示胎盘间充质干细胞可以长期传代到第25代,不同代次的胎盘间充质干细胞增殖能力不同,P5代增殖能力明显高于P10和P15代,临床应用应以P5代之前为宜。
[Abstract]:Background: the proliferation ability of placental mesenchymal stem cells is stronger than that of bone marrow mesenchymal stem cells. But whether the proliferation ability of placental mesenchymal stem cells is the same in every generation is not conclusive. Objective: to analyze the proliferation ability of placental mesenchymal stem cells of different generations. Methods:. Human placental mesenchymal stem cells were isolated by enzyme digestion. Flow cytometry was used to detect stem cell surface markers and adipogenic osteogenesis induced differentiation. Cell count and BrdU method were used to detect cell proliferation ability. Doubling time was calculated according to multiplication time formula. Results and conclusion: placental mesenchymal stem cells were positive for CD440, CD90 and CD105, negative for CD14, CD34 and CD45; After 21 days of adipogenic induction, red fat droplets were observed under the oil red O staining microscope, and calcium salt nodules were observed under the alizarin red staining microscope on 35 days after osteogenesis induction. The number of cells was counted from the first generation. Up to the 15th generation, 3.5 脳 1011 cells. P5 passage was stronger than P10 and P15 generation placental mesenchymal stem cells. When the placental mesenchymal stem cells were subcultured to the 25th passage, the morphology of the 3 specimens was completely changed, showing a paving stone. The other two specimens had almost no adherent cells, suggesting that placental mesenchymal stem cells could be subcultured to the 25th generation for a long time, and the proliferation ability of placental mesenchymal stem cells was different in different generations. The proliferative ability of P5 generation was significantly higher than that of P10 and P15 generation, and the clinical application should be prior to P5 generation.
【作者单位】：吉林大学基础医学院病理学教研室;吉林大学中日联谊医院中心实验室;延边大学药理教研室;吉林省中科生物工程有限公司;
【分类号】：R329
【正文快照】： 文章亮点:1文章比较了胎盘间充质干细胞第5,10,15代次的增殖能力,结果证实细胞增殖能力P5代明显高于P10和P15代。2文章观察了胎盘间充质干细胞连续传代能力和增殖到第15代时产生的细胞数量,并观察和验证了胎盘间充质干细胞传到高代次时细胞形态已发生完全改变的结果。主题词

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