摘要
目的 探讨同种异体大鼠骨髓间充质干细胞在体外的共培养,并行多向分化诱导鉴定。方法 分别通过全骨髓直接培养法及密度梯度法分离培养大鼠骨髓间充质干细胞,各传至第3代后,取等量细胞,混合后共培养,共培养之细胞再次传代后,分别进行成骨细胞、成脂肪细胞诱导,并分别行碱性磷酸酶、Von Cossa及油红染色鉴定。结果 全骨髓直接培养法原代细胞形态不均一,但是增殖能力好;密度梯度法分离得到的细胞较为均一,但增殖力不如全骨髓直接培养法。经过3代传代后,细胞均可获得相对纯化。共培养细胞增殖力较好,细胞形态较为均一,且未出现因免疫排斥导致的细胞死亡。共培养之大鼠骨髓间充质干细胞分别经诱导分化为成骨细胞和脂肪细胞形态（分别经碱性磷酸酶、Von Cossa染色和油红染色证实）。结论 采用全骨髓直接培养及密度梯度离心法,经3-5代培养后均可获得相对纯化之骨髓间充质干细胞。同种异体大鼠骨髓间充质干细胞共培养生长良好,不发生免疫排斥,证实该类细胞的免疫原性极低。在体外,大鼠骨髓间充质干细胞可诱导分化为成骨细胞、脂肪细胞,表明它具有多向分化潜能。
Objectives To investigate the Co-culture of allogenic SD rat bone mesenchymal stem cells （BMSCs） in vitro, and to perform multilineage differentiation potential characterization. Methods Rat BMSCs were isolated and cultured by complete marrow direct culture method and density gradient method respectively, and the 3rd passage cells were harvested and Co-cultured with same con-centration. The passaged cells of the Co-cultured cells were obtained and induced to differentiate into osteoblast and adipocyte respectively, then alkaline phosphatase, Von Cossa and oil red staining characterization were performed. Results The primary cells of complete marrow direct culture method were uneven in shape, but with excellent reproductive activity; the cells obtained by density gradient method are fairly uniform in shape, but with lower reproductive activity; after 3 passages, cells obtained relative purification. The Co-cultured cells were good in reproductive activity and relatively uniform in shape, without cell death because of immunologic rejection. After induction, the Co-cultured cells appeared morphological changes of osteoblast and adipocyte, and were confirmed to be osteoblast by alkaline phosphatase, Von Cossa staining, and be adipocyte by oil red staining. Conclusion Byuse complete marrow direct culture method and density gradient method, after 3-5 passages, relative purified BMSCs can be obtained. The co-cultured allogenic BMSCs are good in growth, without immunologic rejection, imply the weak immunogenicity of BMSCs. In vitro, rat BMSCs can be induced into osteoblasts and adipocytes, which indicates the multilineage differentiation potential of BMSCs.
出处
《苏州大学学报（医学版）》
CAS
北大核心
2007年第2期196-199,206,F0003,共6页
Suzhou University Journal of Medical Science
关键词
同种异体
骨髓间充质干细胞
共培养
鉴定
allogenic
bone mesenchymal stem cells
co-culture
characterization
