大鼠骨髓间充质干细胞诱导分化为脂肪细胞的免疫细胞化学研究

IMMUNOCYTOCHEMICAL STUDY ON INDUCED DIFFERENTIATION OF RAT MESENCHYMAL STEM CELLS INTO ADIPOCYTES

目的 探讨大鼠骨髓间充质干细胞 (mesenchymalstemcells,MSC)分离、纯化和体外诱导分化为脂肪细胞。方法 用密度梯度离心结合贴壁培养、定期换液 ,分离纯化出生大鼠MSC ,传代扩增 ,并用免疫细胞化学法鉴定大鼠MSC的表面抗原。含地塞米松、 3 异丁基 1 甲基黄嘌呤和胰岛素的培养液诱导MSC分化后 ,油红O染色鉴定。结果 大鼠MSC体外扩增 10代以上 ,稳定表达CD4 4、CD5 4、CD10 6。油红O染色显示诱导后 ,71 2 %有脂滴积聚。结论 从大鼠骨髓分离、纯化、体外诱导培养MSC ,可定向分化为脂肪细胞表型。

? Objective To investigate the isolation and purification of rat mesenchymal stem cells (MSC) of bone marrow, and induce them to differentiate into adipocytes in vitro. Method MSCs were isolated from 10-day-old SD rats by gradient centrifugation and anchoring culture. Immunocytochemical stain was adopted to explore the surface antigen of cultured MSCs. After MSCs were induced to differentiate into adipocytes by dexmethason, insulin and isobutylmethylxanthine, Oil Red O stain was employed.Result Rat MSCs were expanded as undifferentiated cells in culture for more than 10 passages and uniformly positive for CD44, CD54 and CD106. After the induction about 71.2% of the MSCs displayedaccumulation of lipid vacuoles,asdetected by oil Red O stain. Conclusion MSCs isolated and cultured from newbornrats express CD44, CD54 and CD106 and can be induced to differentiateinto adipocytes.