中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (10): 1837-1840.doi: 10.3969/j.issn.1673-8225.2011.10.027

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

人羊水间充质干细胞分离培养方法的优化

郭素红1，苑春华2，侯毅鞠1，刘会玲2   

  1. 1吉林医药学院检验系，吉林省吉林市  132013
    2吉林医药学院附属医院妇产科，吉林省吉林市 132013
  • 收稿日期:2010-10-11 修回日期:2010-12-04 出版日期:2011-03-05 发布日期:2011-03-05
  • 作者简介:郭素红★，女，1968年生，吉林省吉林市人，汉族，2003年毕业于哈尔滨医科大学，硕士，副教授，主要从事脑组织损伤及干细胞培养方面的研究。 suhong725@ sina.com
  • 基金资助:

    吉林省教育厅“十一五”科学技术研究项目课题基金资助(吉教科合字:2007-435)。

Optimization of the isolation and cultivation method of human amniotic fluid-derived mesenchymal stem cells

Guo Su-hong1, Yuan Chun-hua2, Hou Yi-jü1, Liu Hui-ling2   

  1. 1Department of Clinical Laboratory, Jilin Medical College, Jilin  132013, Jilin Province, China
    2Department of Gynaecology and Obstetrics, Affiliated Hospital of Jilin Medical College, Jilin  132013, Jilin Province, China
  • Received:2010-10-11 Revised:2010-12-04 Online:2011-03-05 Published:2011-03-05
  • About author:Guo Su-hong★, Master, Associate professor, Department of Clinical Laboratory, Jilin Medical College, Jilin 132013, Jilin Province, China suhong725@sina.com
  • Supported by:

    the Grant for Science Technology Research Program of the Department of Education of Jilin Province during the 11th Five-Year Plan Period, No. 2007-435*

PDF

317

被引次数

4

摘要：

背景：目前关于人羊水间充质干细胞的体外分离培养方法不一，获得较多数量的间充质干细胞仍有困难。
目的：分离80份羊水标本，比较不同的体外培养方法，优化筛选到一种合适的培养纯化条件，为人羊水间充质干细胞的基础研究和临床广泛应用奠定基础。
方法：无菌条件下采集足月分娩和终止妊娠引产的羊水，分离羊水细胞，比较孕期、不同培养基、接种密度、首次换液时间对人羊水间充质干细胞原代培养过程的影响，通过细胞化学染色方法检测细胞代谢情况，观察人羊水间充质干细胞的生物学特性。
结果与结论：相同培养条件下，采用AmnioMAXⅡcomplete羊水专用细胞培养基，以5×104/cm2 的密度接种，首次换液时间为6 d时，人羊水间充质干细胞原代培养成功率较高。孕期为14~20周的人羊水间充质干细胞培养成功率高于晚期妊娠(21~36周)及足月分娩(37~42周)的羊水。人羊水间充质干细胞强表达过碘酸-雪夫、酸性磷酸酶、酸性α-醋酸萘酚酯酶染色，弱表达中性粒细胞碱性磷酸酶，不表达过氧化酶、苏丹黑染色。提示，优化筛选到一种合适的人羊水间充质干细胞培养纯化条件。

关键词: 羊水, 间充质干细胞, 细胞培养, 优化, 足月分娩

Abstract:

BACKGROUND: There are different methods of in vitro isolating culture for human amniotic fluid-derived mesenchymal stem cells (AF-MSCs). To gain most AF-MSCs is still difficult.
OBJECTIVE: We had isolated 80 cases AF, and compared different in vitro culture methods for optimization and bolted a better method to establish foundation of AF-MSCs in clinical widespread application.
METHODS: Amniotic fluid was collected from full-term deliveries and artificial labors under aseptic condition. The amniotic fluid cells were isolated by centrifugation. The influences of different pregnancy ages, culture media, incubation densities and the time points of the first medium change on the growth of AF-MSCs were analyzed. The cell metabolism was determined and biological characteristics of AF-MSCs were observed by cytochemical staining.
RESULTS AND CONCLUSION: When the other conditions were the same, AmnioMAXⅡcomplete medium was the best medium. 5×104/cm2 was the best planting density; the best time for the first medium change was on the sixth day. The achievement ratio of amniotic fluid from fetation of intermediate stage (14-20 weeks) was higher than that of later fetation (21-36 weeks) and full-term deliveries (37-42 weeks) when the other conditions were the same. AF-MSCs highly expressed periodic acid-schiff, acid phosphatase, α-naphthol acetate esterase (α-NAE) and weakly expressed neutrophil alkaline phosphatase, but no peroxidase and Sudan black was found. Results suggest that we have found the optimized method of culturing AF-MSCs.

中图分类号: