人骨髓间质干细胞库的初步建立

目的 探讨建立生物学性状稳定的人骨髓间质干细胞(human marrow mesenchymal stem cells，hMSCs)库的可行性，为组织工程种子细胞的来源作初步探索。方法对分离得到的hMSCs采用液氮低温冻存，并在一定时间复苏培养，以流式细胞仪检测其表面抗原，在诱导培养基中对其行成骨和成软骨诱导培养，光镜：电镜观察细胞的形态，采用组织化学、免疫组织化学及分子生物学的方法检测成骨和成软骨诱导的特异标志物：碱性磷酸酶、Ⅰ型胶原、Ⅱ型胶原及骨钙素，研究其生物学性状。结果从骨髓中分离得到了纯度达96％以上的hMSCs，复苏后的hMSCs形态和表面抗原保持不变，可继续扩增10代以上，倍增时间40h。复苏后第2、6、10代hMSCs均保持较强的向软骨细胞、成骨细胞的分化能力。结论初步建立了hMSCs库，可为骨及软骨组织工程提供较好的种子细胞。

INCIPIENT ESTABLISHMENT OF HUMAN BONE MARROW MESENCHYMAL STEM CELLS BANK

OBJECTIVE: To investigate the possibility of establishing the human bone marrow mesenchymal stem cells (hMSCs) bank as to provide an alternative source for the seed cells of tissue engineering. METHODS: The cell surface antigens of the purified, expanded hMSCs and the ones following cryopreservation were detected by flow cytometry, cultured in a special medium to induce the ostoegenic and chondrocytic differentiation. Morphology was studied by light and electronic microscopes. The detection of alkaline phosphatase, collagen type I, osteocalcin, and collagen type II were also performed by immunochemistry and molecular biology. RESULTS: The phenotype and expansion possibility of hMSCs after cryopreservation were remained. It could expand for 10 generations. The doubling time was 40 h. CONCLUSION: The bank of hMSCs is incipiently established and can provide eligible seed cells for tissue engineering.