4种来源的间充质干细胞的生物学特性比较

Comparison of biological characteristics of mesenchymal stem cells derived from four kinds of tissues in rabbit

目的比较骨髓、骨骼肌、肋软骨膜及软骨来源的间充质干细胞(mesenchymal stem cells,MSCs)的生长、免疫表型及体外诱导分化能力的差异。方法分别获取新西兰大白兔的骨髓、骨骼肌、肋软骨膜细胞及软骨细胞,体外培养观察其生长形态、表型特征及其诱导成骨、成脂的分化能力。结果 4种来源的MSCs均从第2代开始转变为梭形,第3代逐渐出现漩涡状排列;骨髓、骨骼肌和软骨膜来源的细胞生长状态良好,可传10代以上,但软骨来源的细胞在第5代后,逐渐退变死亡。骨髓来源的原代细胞部分表达CD29、CD90、CD34,传代后不表达CD34,但CD29、CD90的表达逐代增强;骨骼肌、软骨膜及软骨来源的原代细胞均不表达CD29、CD90和CD34,从第2代开始均表达CD29、CD90,且不断增强;仅原代软骨细胞表达Ⅱ型胶原,至第2代基本不表达;无细胞表达Desmin。4种来源的第3代细胞经诱导均可分化为成骨细胞或成脂细胞。结论 4种来源的干细胞均具有MSCs的特点,但骨髓、骨骼肌、软骨膜来源的干细胞的生长明显强于软骨来源的干细胞。

Objective To compare the growths,immunophenotypes and abilities in induction of differentiation in vitro of mesenchymal stem cells（MSCs）derived from four kinds of rabbit tissues.Methods Cells were isolated from the bone marrow,skeleton muscle,rib perichondrium and rib cartilage of New Zealand white rabbits,cultured in vitro,and observed for growth,phenotype and ability in induction of osteogenic and adipogenic differentiations.Results The cells derived from four kinds of tissues were in a spindle shape starting from passage 2,of which the passage 3 were in a whirlpool arrangement.The cells derived from bone marrow,skeleton muscle and rib perichondrium grew well and could be subcultured for more than ten passages.However,senescence and death were observed in the cells derived from rib cartilage after subculture for more than five passages.CD29,CD90 and CD34 were expressed in primary cells derived from bone marrow.In these cells after subculture,no CD34 was expressed,while the expression levels of CD24 and CD90 increased with the increasing passage.However,no CD29,CD90 or CD34 was expressed in primary cells derived from skeleton muscle,rib perichondrium and rib cartilage,while CD29 and CD90 were expressed in the cells after subculture for more than two passages,and the expression level increased with the increasing passage.Collagen Ⅱ was only expressed in primary cells derived from rib cartilage but disappeared in the cells of passage 2.Desmin was unexpressed in any cells.All of cells of passage 3 derived from four kinds of tissues could be differentiated into osteoblasts or adipose cells.Conclusion All of the cells derived from four kinds of tissues showed characteristics of MSCs.However,the growths of stem cells derived from bone marrow,muscular tissues and rib perichondrium were superior to that derived from rib cartilage.