人胎儿骨髓间充质干细胞的分离及生物学鉴定

通过原代细胞培养,从引产胎儿骨髓组织中分离干细胞,然后进行生物学鉴定,旨在体外建立培养胎儿骨髓干细胞的有效方法,为进一步研究干细胞奠定基础．本研究对四个月的引产胎儿骨髓组织进行原代细胞培养,采用贴壁筛选法,在含有15％胎牛血清的L-DMEM／IMDM(1:1)混和培养液中培养,7 d后细胞可长满瓶底．显微镜下观察,细胞形态均一,呈长梭形．传代后,在8代以内的细胞贴壁能力较强,生长速度较快．将其命名为BMMS-03．在第3代时,对培养的细胞进行了于细胞标志物的生物学鉴定,采用流式细胞仪对经免疫荧光染色的细胞进行检测．结果显示:97．2％的细胞呈CD105阳性反应,66．0％的细胞呈CD106阳性反应, 9．2％的细胞呈CD34阳性反应．阴性对照组阳性反应为0．5％．生物学鉴定的初步结果提示,从胎儿骨髓组织中分离培养成功的细胞为骨髓间充质干细胞,其细胞形态学特征、CD105 、CD106 和CD34-的检测结果均符合间充质干细胞的特征．本研究成功地建立了体外培养胎儿骨髓间充质干细胞的有效方法,所获得的间充质干细胞纯度较高,增殖较快,适用于干细胞生物学和组织工程学的研究．

Isolation and Identification of Mesenchyal Stem Cells Derived from Human Fetal Bone Maroow

In order to establish an effective method of isolating stem cells and lay a good foundation for the study, the stem cells were isolated from elicited fetal bone marrow tissue by primary cell culture. In this study the stem cells from bone marrow of a four-month old elicited fetus were primarily cultured in the mixed medium of L-DMEM/IMDM (1 : 1) containing 15 % fetal bovine serum (FBS) by the method of adhering to culture plastic. The cells covered the bottle bottom after 7 days. The morphology of the cells was homogeneous in fiber-like spindle shape. The cells within eight generations grew up well. The biological characteristics of the cells from the third generation were immunofluorescently identified by flow cytometry analysis. The results showed that the positive rate for CD105 was 97. 2 %, 66. 0 % for CD106 and 9. 2 % for CD34. The positive rate was 0. 5 % in negative control cells. The primary identification demonstrated that the cells derived from fetal bone marrow were mesenchymal stem cells (MSCs) according to the morphology of MSCs and the markers of stem cells, such as CD105 , CD106 and CD34~ In this text the effective method of isolating MSCs from fetal bone marrow was established, and the cells were homogeneous and grew up quickly. The isolated MSCs provide the materials for studying stem cells and tissue engineering.