骨髓胚胎样干细胞的形态结构特征观察  

A study on phenotype and ultramicrostructure of embryonic-like stem cell derived from adult bone marrow

作　　者：庞荣清[1] 张永云[1,2] 邓永丽[1] 马丽花[1] 何洁[1] 赵晶[1] 潘兴华[1] 张成[3] 

机构地区：[1]成都军区昆明总医院干细胞与组织器官工程研究中心,云南昆明650032 [2]云南农业大学基础实验研究中心,云南昆明650201 [3]中山大学附属第一医院神经内科,广东广州510080

出　　处：《中国实验诊断学》2012年第9期1608-1611,共4页Chinese Journal of Laboratory Diagnosis

基　　金：云南省自然科学基金面上项目(NO.2009ZC151M)

摘　　要：目的观察胚胎样干细胞(ELSCs)的形态结构特征。方法分别采用无血清培养基在明胶包被过的培养瓶和传统方法在未包被的培养瓶中分离骨髓胚胎样干细胞和间充质干细胞(MSCs),以MSCs为对照,在光学显微镜下观察ELSCs的细胞形态,在透射电镜下观察ELSCs的超微结构,并采用染色体技术分析传代细胞的核型。结果与MSCs相比,ELSCs体积更小、形态纤细均一,传代后细胞贴壁快,在细胞培养瓶中均匀分布生长,不易老化。第四代ELSCs的绝大多数细胞膜显示显著的花多样伪足,细胞浆中有丰富的粗面内质网。而部分MSCs的细胞膜光滑,部分MSCs膜表面有微绒毛,多数细胞胞浆内有空泡和丰富的粗面内质网而且明显扩张形成内质网池,内含中等电子密度的分泌物。第十代ELSCs和MSCs都有正常的核型。结论 ELSCs具有比MSCs更不成熟的细胞形态结构特征,传代10代后细胞仍然是安全的。Objective To investigate morphological and structural characters of embryonic-like stem cells （ELSCs） from adult bone marrow. Methods ELSCs were isolated from bone marrow by culturing in flask coated-gelatin with the serum-free medium. Compared to mesenchymal stem cells （MSCs） isolated from the same marrow by traditional meth- od. Morphous of ELSCs were observed under light microscope,and ultramicrostructure of ELSCs were observed under transmission electron microscope,and karyotype of passaged ELSCs was analyzed by chromosome technique. Results Compared to MSCs, ELSCs appeared as small, morphologically slenderer and homogeneous, and after passage, they were quickly adherent to flask coated-gelatin and grew well-distributed in the flask,and were not more readily ageing. Under transmission electron microscope, Most of passage 4 ELSCs revealed prominent flower-like pseudopodium on its cell membrane,abundant rough endoplasmie retieulum profile in its cytoplasm. As a control,some passage 4 MSCs revealed a smooth cell membrane and another cells revealed microvilli on its cell membrane. Major MSCs showed distinctive vac- uoles and significant rough endoplasmic reticulum cisternae with moderately electron-dense secretory material. Chromosome analysis demonstrated the karyotypes of passage 10 both ELSCs and MSCs were normal state. Conclusion ELSCs showed more immature morphological and structural characters than MSCs,passage 10 cells have been safe.

关 键 词：多潜能干细胞 骨髓 超微结构 细胞形态 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]