中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (14): 2525-2529.doi: 10.3969/j.issn.1673-8225.2011.14.013

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

马血清在脂肪干细胞成肌诱导中的作用

刘  杰，傅  强   

  1. 上海市第六人民医院，上海市  200233
  • 收稿日期:2010-10-29 修回日期:2010-12-07 出版日期:2011-04-02 发布日期:2013-11-02
  • 通讯作者: 傅强，博士，主任医师，上海市第六人民医院，上海市200233
  • 作者简介:刘杰★，男，1984年生，山东省滨州市人，汉族，苏州大学在读硕士，主要从事组织工程化尿道研究。 shaliujie@126.com

Horse serum effects on the differentiation of adipose-derived stem cells into myoblasts

Liu Jie, Fu Qiang   

  1. Shanghai Sixth People’s Hospital, Shanghai  200233, China
  • Received:2010-10-29 Revised:2010-12-07 Online:2011-04-02 Published:2013-11-02
  • Contact: Fu Qiang, Doctor, Chief physician, Shanghai Sixth People’s Hospital, Shanghai 200233, China
  • About author:Liu Jie★, Studying for master’s degree, Shanghai Sixth People’s Hospital, Shanghai 200233, China

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340

被引次数

1

摘要：

背景：利用5-氮杂胞苷诱导脂肪干细胞成肌实验对培养技术以及细胞活性要求较高，诱导成肌时间较长。倘若某种添加剂可缩短诱导时间，将具有重要意义。
目的：评估马血清在5-氮杂胞苷诱导脂肪干细胞成肌细胞实验中的作用。
方法：将第1代长满100 mm培养皿的脂肪干细胞传代至3个6孔板，实验组培养基中加入体积分数为5%马血清+10 μmol/L 5-氮杂胞苷；对照组培养基中单纯添加10 μmol/L的5-氮杂胞苷；空白组为单纯低糖DMEM培养基，余培养、传代及鉴定所需的条件相同。每日光镜观察记录形态，在培养的第7，14，28，35天行免疫荧光和流式细胞仪检测肌蛋白表达差异。
结果与结论：免疫荧光检测成肌特异性胞质蛋白表达以及流式细胞检测相应蛋白表达率提示，加入马血清后的实验组脂肪干细胞的成肌速度和诱导成肌所需时间明显比单纯加入5-氮杂胞苷的对照组更有优势。实验初步表明马血清在促进脂肪干细胞诱导成肌方面起着缩短诱导成肌所需时间的作用。

关键词: 马血清, 脂肪干细胞, 成肌诱导, 5-氮杂胞苷, 成肌特异性胞质蛋白

Abstract:

BACKGROUND: 5-azacytidine (5-aza)-induced adipose-derived stem cells (ADSCs) into myoblasts required advanced techniques and high cell viability, and long induction time. If certain additive can shorten induction time, this additive would be significant.
OBJECTIVE: To assess the horse serum effects on 5-aza-induced ADSCs into myoblasts.
METHODS: The first generation of ADSCs fully covered the 100-mm culture dish was passaged to three 6-well culture dishes. In the experimental group, 5% horse serum + 10 μmol/L 5-aza was added in the culture medium. In the control group, 10 μmol/L of 5-aza was simply added. In the blank group, low glucose DMEM medium alone was used; the necessary conditions of culture, subculture and identification are the same. Morphology was observed daily by light microscopy. Immunofluorescence and flow cytometry were used to test the muscle protein differences on days 7, 14, 28, 35.
RESULTS AND CONCLUSION: Immunofluorescence and flow cytometry detection for the expression of corresponding muscle-specific cytoplasmic protein showed that time of inducing into myoblasts in the experimental group was shorter than the control group. Studies primarily suggested that the horse serum can shorten the time in 5-aza inducing ADSCs into myoblasts.

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