小鼠骨髓基质细胞成内皮细胞分化及其缺血区存活能力研究  

Endothelial differentiation and survival ability of mouse bone marrow-derived stromal cells.

作　　者：张坡[1] 黄岚[1] 刘艳霞[2] 朱鲜阳[2] 宋明宝[1] 于世勇[1] 

机构地区：[1]第三军医大学新桥医院全军心血管内科中心,重庆400037 [2]沈阳军区总医院,辽宁沈阳110016

出　　处：《中国实用内科杂志：临床前沿版》2006年第9期1404-1406,共3页

基　　金：国家自然科学基金项目(30270568)

摘　　要：目的 探讨体外培养的骨髓基质细胞的部分生物学特性、成内皮细胞分化能力及在缺血区存活能力？方法 2005年1月至11月，第三军医大学新桥医院全军心血管内科中心分离5～6周龄的小鼠胫骨、股骨，预冷的DMEM／F12培养基冲洗出骨髓，密度梯度离心分离出骨髓单核细胞，接种后12～16d形成单层贴壁的成纤维样细胞：体外诱导分化鉴定分离的细胞：建立下肢缺血模型，荧光标记的体外扩增的骨髓基质细胞被移植入缺血组织，移植后1周，荧光显微镜及HE染色，检查荧光强弱分布与HE染色密度的时空关系。结果 体外传代培养的基质细胞诱导后分泌NO，移植1周在缺血区检测到大量荧光阳性细胞存活。结论在本实验条件下，体外培养的骨髓基质细胞生长稳定，传代后的细胞增殖较快，表现出较早期细胞特点，在体外能分化为血管内皮细胞，可望用作缺血性心脏病的细胞治疗的供体细胞。Objective To studv some biological properties of mouse bone marrow stromal cells （MSCs） and their ability to differentiate endothelial cells and surviving ability in ischemic tissue, and provide an experimental foundation for applying MSCs to ischemic repair. Methods After the tibias and femurs were dissected from 5 - to 6 - week - old mice from Jan. 2005 to Nov. 2005, the marrow was flushed out with ice - cold DMEM/F12 medium. The mononuclear cells of the marrow were obtained with density gradient centrifugation and the plated and cultured in DMEM/F12 medium. The cultured cells in vitro were induced with endothelial cell growth supplement. The ability of MSCs was also was examined in ischemic tissue. Results The adherent fibroblast - shaped cells approached coniluence in single layer 12 - 16 d after plating. The cultured MSCs in vitro differentiated into endothelium. Numerous scattered DAPI - labeled cells were found in the specimen seven days after implantation, and bematoxylin and eosin staining of the adjacent section showed concordance between dense hematoxylin staining and presence of DAPI epifluorescence, and there was no obvious inflamma- tory＂ response. Conclusion The subcultured MSCs possess potential to differentiate into endothelial cells. MSCs show stable growth in vitro, easy survival in the subculture and rapid proliferation in present culture condition. MSCs may be used in therapy for myocardium ischemia.

关 键 词：骨髓细胞 细胞培养 分化 缺血 存活 

分 类 号：R5[医药卫生—内科学]