外周血作为骨组织工程种子细胞来源的初步研究

A pilot study on the peripheral blood as a source of seed cells for bone tissue engineering

目的:研究外周血作为骨组织工程种子细胞来源的可能性。方法:密度离心法分离兔外周血单个核细胞,冲洗收集长骨内骨髓,将两者在体外进行培养。检测两种来源细胞的表面分子标志物及成脂、成骨能力差异。结果:骨髓原代培养细胞集落形成早,细胞形态均一。传代后CD29阳性,CD34阴性;成脂诱导时脂滴出现早且多;成骨诱导时碱性磷酸酶、Ⅰ型胶原表达早,矿化结节数量多且体积大。外周血原代培养细胞集落形成晚,形态不一,种类多。传代后CD29阳性,CD34弱阳性;成脂诱导时脂滴出现晚且少;成骨诱导时碱性磷酸酶、Ⅰ型胶原表达较晚,有散在的矿化结节和管样结构形成。结论:外周血内含有间充质干细胞、内皮祖细胞等多种干细胞,可以作为血管化组织工程骨的种子细胞来源,但需要进一步优化培养方法。

Objective： To investigate whether peripheral blood could act as a source of seed ceils for bone tissue engineering. Methods： Mononucleated cells in rabbit peripheral blood were separated by density centrifugalization. Bone marrow in long bones was collected. The samples were cultured in vitro. Cell markers, lipogenic and osteogenic ability of cells from these two sources were compared. Results： Cell colonies of bone marrow formed early with uniform fibroblast-like cells. These cells were positive of CD29 and negtive of CD34. Lipid droplets formed early and markedly when adipo-induced. Alkaline phosphatase and type I collagen expressed early and bulk calcium nodes formed. Cell colonies of peripheral blood formed late with different kinds of morphilogical cells. These cells were positive of CD29 and weakly positive of CD34. Lipid droplets formed late and partly when adipo-induced. When osteo-induced, alkaline phosphatase and type I collagen expressed late and less, small calcium nodes formed with tubule-like structures here and there. Conclusion： Peripheral blood contained some kinds of stem cells including mesenchymal stem cells and endothelial progenitor cells. It was a possible source of seed cells for vascularized bone tissue engineering. But improved culture procedures should be studied in the future.