脂肪间充质干细胞的基本生物学特性及向成骨细胞诱导分化的实验研究

Biological characteristics and osteoblast differentiation of adipose mesenchymal stem cells

目的 研究脂肪间充质干细胞的基本生物学特性以及在特定培养条件下向成骨细胞分化 ,探讨其作为骨组织工程的种子细胞的可行性。方法 取 3周龄Lewis大鼠的腹股沟脂肪垫 ,消化法获得脂肪间充质干细胞 ,分别用脂肪诱导培养基和成骨诱导培养基诱导其向脂肪细胞与成骨细胞分化 ,组织化学染色、免疫细胞化学染色和Westernblot检测细胞分化的情况。结果 从成体大鼠脂肪组织中培养出脂肪间充质干细胞 ,原代脂肪间充质干细胞能自发分化为脂肪细胞 ,传代细胞在胰岛素和呋塞米的作用下生成脂滴 ,过氧化物酶体增殖物激活受体 (PPAR)γ表达增强 ,向脂肪细胞分化 ;在呋塞米、抗坏血酸、β 甘油磷酸钠的诱导下 ,脂肪间充质干细胞的碱性磷酸酶(ALP)活性检测显示诱导组与对照组差异有显著性 (P <0 .0 1) ,vonKossa染色出现钙结节 ,骨桥蛋白 (OPN)、骨形态发生蛋白 (BMP) 2免疫细胞化学染色阳性 ,Westernblot检测到诱导后细胞OPN、BMP2的表达。结论 从脂肪组织中可获得具有多分化潜能的间充质干细胞 ,经诱导后可分化为脂肪细胞和成骨细胞 。

Objective To investigate the basic biological characteristics and osteoblast differentiation of adipose mesenchymal stem cells as seeding cells in bone tissue engineering.Methods Adipose mesenchymal stem cells were obtained from inguinal fat pads of Lewis rat with digestion.Adipose mesenchymal stem cells were induced into adipocytes and osteoblasts by different revulsant,and then the differentiations were examinedby cytochemistry,immuncytochemistry and Western blotting.Results A population of adipose mesenchymal stem cells were successfully isolated from rat adipose tissue.Primary cells could differentiated into adipocytes spontaneously,and passaging cells must be induced to differentiate into adipocytes with insulin and dexamethasone.After treated with dexamethasone,ascorbic acid,and beta sodium glycerophosphate,the differentiation of adipose mesenchymal stem cells towards osteoblasts was verified by the positive results of ALP,OPN and BMP2 through cytochemistry,immuncytochemistry and Western blotting respectively.Conclusion Mesenchymal stem cells with multilineage potential obtained from rat adipose tissue can differentiate to adipocytes and osteoblasts when cultured in vitro,and adipose mesenchymal stem cells can likely be served as optimal autogenous cell source for bone tissue engineering.