兔骨髓基质干细胞培养及鉴定

目的：对新西兰大白兔骨髓基质干细胞（BMSCs）进行体外培养及扩增，观察其原代及传代细胞的生长特点及生物学特点，获得足够量的细胞用于材料细胞相容性实验。方法：骨髓髓腔冲洗获得新西兰大白兔幼兔股骨骨髓，用全骨髓培养法进行体外培养获得BMSCs，胰酶消化传代，用条件培养基培养传代细胞。逐日倒置显微镜观察细胞生长情况，对传代细胞进行HE染色和碱性磷酸酶（ALP）染色。结果：幼兔骨髓基质干细胞体外培养生长良好，原代细胞约2周汇成单层，传代后1周左右长满瓶底。HE染色光镜下观察见兔骨髓基质干细胞为单核细胞，细胞呈梭形或不规则长多边形，传代细胞碱性磷酸酶染色呈强阳性。结论：兔骨髓基质干细胞的体外增殖能力强，可诱导为成骨细胞，可作为骨缺损材料细胞相容性研究的受试细胞。并且可以作为骨组织工程的种子细胞。

Culture and identification of the rabbit bone marrow stromal cells in vitro

Objective:To expore the biological features and the proliferation of primary and passaged cells of bone marrow stromal cells(BMSCs) of rabbit cultured in vitro,and we can acquire enough BMSCs to pursue our study on the material's cell histocompatibility with rabbit bone marrow stromal cells.Method:Bone marrow was harvested from rabbit femur bone marrow with flushing method..Adherent cells were selected as BMSCs after the whole marrow was cultured.Trypsin was used to digest and passage.Passaged cells were fed with conditioned medium,and observed under inverted phase contrast microscope every day.Passaged cells were examined by HE staining and histochemistry staining for ALP.Result: The BMSCs of rabbit cultured grew and survived well in vitro.The primary BMSCs were confluent in about two weeks,and passage cells took less than one week to cover the bottle bottom.Observed by HE staining,BMSCs became monocytes,in the spindle or long multiangular shape.Passaged cells showed strong positive staining of Alkaline phosphatase.Conclusion: Since the BMSCs of rabbit cultured can proliferate well and can be induced to differentiate into osteoblasts in vitro,they can be used as test cells to evaluate the cell histocompatibility of bone defect substitution materials.They can also be used as seed cells for bone tissue engineering.