不同血清培养条件下骨髓间充质干细胞的增殖

背景：目前，分离培养骨髓间充质干细胞应用最为广泛的细胞生长添加剂是胎牛血清/小牛血清，应用脐血清进行骨髓间充质干细胞培养的相关研究较少。 目的：观察不同血清培养对骨髓间充质干细胞增殖的影响。 方法：用Ficoll密度梯度离心法分离纯化骨髓间充质干细胞，在含脐血清、胎牛血清以及无血清的L-DMEM培养基中培养，观察各组细胞形态、增殖情况；应用流式细胞术对细胞表面抗原CD34、CD44、CD45、CD105进行表型鉴定。 结果与结论：脐血清培养后细胞形态较小，似纺锤状，细胞呈网状生长；胎牛血清培养后细胞呈梭形成纤维细胞样，呈集落样生长；脐血清和胎牛血清均能促进细胞生长增殖，但骨髓间充质干细胞在脐血清中增殖更活跃，无血清培养基本无增殖。流式细胞鉴定显示CD34、CD45阴性，CD44、CD105阳性，脐血清扩增后骨髓间充质干细胞的表面标记无明显改变。

Proliferation of bone marrow mesenchymal stem cells under different serum culture conditions

BACKGROUND: There are many studies concerning bone marrow mesenchymal stem cells for culturing methods, and fetal bovine serum (FBS)/ fetal calf serum (FCS) are used widely. However, there is few study about cord blood serum (CBS) culture method on bone marrow mesenchymal stem cells (BMSCs). OBJECTIVE: To explore the effects of different serum culture method on the proliferation of BMSCs. METHODS: Ficoll density gradient centrifugation method was used to isolate and purify BMSCs that were incubated in L-DMEM containing CBS and FBS and serum-free media. Morphology and proliferation of BMSCs were observed. CD34, CD44, CD45 and CD105 were identified using flow cytometry. RESULTS AND CONCLUSION: CBS group cells appeared smaller, more spindle-shaped with fewer cytoplasmic processes than the cells cultured with FBS. CBS and FBS could promote cell growth and proliferation, but BMSCs in CBS were active, but no proliferation was determined in serum-free media. Flow cytometry showed that BMSCs were negative for CD34, CD45, but positive for CD105 and CD44. Surface marker on BMSCs did not significantly change following CBS amplification.