摘要
本研究目的是分离富集小鼠骨髓间充质干细胞(mMSC),鉴定其生物学特性和多向分化潜能。取4-5周龄雄性C57BL/6小鼠骨髓细胞,采用全骨髓贴壁法和单克隆培养法分离、纯化和扩增mMSC;分析细胞免疫表型、生长曲线、细胞周期;进行多向分化潜能鉴定;传代培养达30代后,进行成瘤性检测。结果表明:建立的小鼠间充质干细胞系在体外可连续传代培养达30代,细胞仍保持多向分化潜能,细胞高表达CD29、CD44、Sca-1、MHC-Ⅰ,中度表达CD13、CD90.2,不表达CD117、CD45、Flk-1、MHC-Ⅱ类抗原。mMSC体外能诱导分化成骨、脂肪、软骨细胞。结论:全骨髓贴壁法和集落培养法富集可获得mMSC,其在体外连续传代30代以上仍能维持生物学特性稳定,具有高度的增殖和多向分化潜能,无成瘤性。
The study was aimed to isolate and establish mesenchymal stem cell line from adult murine bone marrow as well as to identify its biological characteristics and differentiation potential. Bone marrow cells （BMCs） were collected by flushing the femurs and tibias of 4-5-week-old male C57BL/6 mice, and were inoculated at a concentration of 1 × 10^6/cm^2. mMSCs were isolated, enriched and expanded by using bone marrow adherant culture and monoclonal culture. The characteristics of the cells, such as morphology, growth pattern, cell cycle, phenotype, karyotype and multipotent differentiation potential, cytogenetic stability and tumorigenesis were determined. The results indicated that the cell population consisted of spindle- and star-shaped cells, they were highly positive for CD29, CD44, Sea-1, MHC- Ⅰ , moderate positive for CD13, CD90.2 and negative for CDll7, CD45, Flk-1 and MHC-Ⅱ. mMSCs could be induced to differentiate into adipocytes, osteoblast cells and chondrocytes. It is concluded that mMSC can be isolted, expanded and enriched by using bone marrow adherent culture and monoclonal culture. No tumor formations are observed for 3 months in nude mice after subcutaneous injection, mMSCs retain their properties after at least 30 passages in culture as well as from frozen stocks.
出处
《中国实验血液学杂志》
CAS
CSCD
2007年第3期542-546,共5页
Journal of Experimental Hematology
基金
国家"863"高技术发展项目(编号2002AA205051)
上海市科委重大基础研究基金资助项目(编号03DJ14020)
上海市科委科技攻关项目(编号05DZ19327)
关键词
间充质干细胞
骨髓
细胞培养
生物学特性
mesenchymal stem cell
bone marrow
cell culeure
biological characteristics
