摘要
目的:探讨成年大鼠的骨髓间充质干细胞(MSCs)的分离、体外培养方法,为其应用提供大量种子细胞。方法:用密度梯度离心结合贴壁培养法分离纯化大鼠骨髓MSCs,传代扩增,对细胞进行免疫组化染色鉴定。结果:取得较高纯度的成体大鼠骨髓MSCs,并保持细胞的活性;成体大鼠骨髓MSCs在体外培养中为贴壁生长,培养3～4d后开始大量增殖,并形成星形、梭形的细胞增殖集群,对分离后所获得的细胞进行免疫组化染色,细胞CD44,FN均阳性,CD34,CD31均阴性。结论:采用密度梯度离心结合贴壁培养法可获得较高纯度的MSCs,是实用、便捷和可行的方法。
AIM： To explore a practical method of separation and culture in vitro of mesenchymal stem cells （MSCs） from bone marrow of the adult rats, and to offer plenty of seed cells for its actual application.
METHODS： MSCs from adult rat bone marrow were separated and expanded by gradient centrifugation and adherence culture. Then the cells were identified by immunohistochemistry method.
RESULTS： MSCs separated from the bone marrow of the adult rats were collected in higher purity by means of gradient centrifugation. These cultured cells grew in an adherent mode on the wall. They were able to greatly proliferate after 3-4d culturation with shape of asteroid and fusiform. The results of immunohistochemistry of the cultured cells showed that both CD44 and FN were positive and CD34 and CD31 were negative.
CONCLUSION： It is a simple and practical method to separate MSCs from the bone marrow of adult rats by means of gradient centrifugation and adherence to the culture plastic. MSCs can greatly proliferate in vitro and will be very useful for its further research and application.
出处
《国际眼科杂志》
CAS
2006年第3期562-564,共3页
International Eye Science
基金
中国国家自然科学基金资助项目(No.30371515)~~
关键词
间充质干细胞
细胞培养
骨髓
大鼠
mesenchymal stem cells
cell culture
bone marrow
rats
