来源于脂肪组织的基质细胞向成骨细胞分化

Human adipose tissue-derived stromal cells differentiate into osteogenic cells

目的 :研究来源于脂肪组织的基质细胞体外培养和向成骨细胞分化条件。方法 :常规方法培养脂肪组织来源的基质细胞 ,向成骨细胞分化诱导 ,应用免疫细胞化学方法对细胞进行鉴定 ,碱性磷酸酶法对分化的成骨细胞鉴定。结果 :从成年人的脂肪组织中分离出基质细胞 ,在体外生长形态类似成纤维细胞 ,可以维持在未分化状态稳定增殖 ,体外可持续扩增和传代。在一定的条件下可诱导分化为成骨细胞 ,分化的细胞表达碱性磷酸酶和Ⅰ型胶原 ,在培养皿中也发现钙化斑。结论 :脂肪组织中存在的基质细胞能分化为成骨细胞 。

Objective: To investigate the optimal culture conditio ns for adipose tissue-derived stromal cells(ADSCs) and for the induction of these cells to differentiate into osteogenic cells. Methods: ADSCs were cultured with routine methods,bFGF at 20 ng/ml was added into the medium and the proliferative of ADSCs was examined by cell counting. 0.1 μmol /L of dexamethasone,10 mmol/L of β-glycerophosphate and 50 μmol/L of ascorbic acid were adapted to induce the cells to differentiate into osteogenic cells, ADSCs were identified by immunocytochemistry and differentiated osteogenic cells were identified by alkaline phosphatase(AP) staining and immunocytochemistry. Result: A population of ADSCs could be isolated from adul t human adipose tissue,the cells were fibroblast-like and could be maintaine d in vitro for extended periods with stable population doubling.The cells w ere expanded as undifferentiated in culture for more than 10 passages, indicati ng their proliferative capacity.bFGF stimulated the cell proliferation.Dexameth asone,β-glycerophosphate and ascorbic acid induced (40±8.6)% of ADSCs to ex press alkaline phosphatase(AP) ,(35±10.6)% of AP positive ADSCs were found to be collagen I positive. Calcification plaques were occasionally found in the cul tures. Conclusion:The data support the hypothesis that adu lt human adipose tissue contains stem cells capable of diffferentiating into ost eogenic cells.