大鼠胎盘来源间充质干细胞的生物学特性

Biological characteristics of rat placenta-derived mesenchymal stem cells

背景:研究报道,人间充质干细胞用于实验动物研究的结果不令人满意,所以考虑使用大鼠胎盘来源的间充质干细胞用于动物实验研究,以期获得更好的实验结果。查阅国内外相关文献,目前尚未发现大鼠胎盘来源间充质干细胞的相关报道。目的:建立从大鼠胎盘分离间充质干细胞的方法,观察其基本生物学特性。方法:通过胶原酶消化法从大鼠胎盘分离并得到间充质干细胞。每天用倒置显微镜观察其形态。用MTT法测定其生长动力学并绘制生长曲线。用流式细胞仪检测其表型及细胞周期。通过免疫组化法证实其成脂及成骨分化的潜能。结果与结论:原代细胞培养8h后细胞贴壁,24h内细胞形成集落,第4代细胞大小、形态基本一致,以梭形为主。细胞周期检测提示G0/G1期、S期、G2期的比例分别为83.76%,8.01%,8.23%。免疫表型分析其表达CD29和CD90,不表达CD45。体外诱导实验证实胎盘间充质干细胞具有成脂和成骨分化的潜能。

BACKGROUND：Mesenchymal stem cells（MSCs） have the potential of multilineage differentiation. Some researchers put them into tissue-regeneration or diseases treatment. There are many studies about MSCs derived from bone marrow,umbilical cord of rat or mouse. However,little is known about MSCs derived from rat placenta. OBJECTIVE：To establish a method to isolate MSCs from rat placenta and to observe its biological characteristics in vitro. METHODS：Rat placental MSCs（r-pl-MSCs） were separated and obtained by collagenase-digested method. Morphology of r-pl-MSCs was daily observed with inverted microscope. Growth kinetics was measured with MTT assay and growth curve was drawn. Their surface antigens and cell cycle were detected by flow cytometer. Adipogenic differentiation and osteogenic differentiation were tested by immunohistochemistry. RESULTS AND CONCLUSION：The primary cultured cells adhered after 8 hours and formed clone within 24 hours. The size and morphology of the 4th generation cells were almost same;most cells were fusiform-shaped. Cell cycle measure showed the proportion of cells in G0/G1 phase,S phase,G2 phase was respectively 83.76%,8.01%,8.23%. Immunophenotype analysis showed that the cells expressed CD29 and CD90,but no CD45. Placenta-derived MSCs have potentials of adipogenic and osteogenic differentiation in special culture condition in vitro.