提高培养内皮祖细胞成功率的探讨

摘要

Objective To study an optimal culture condition for rat endothelial progenitor cells (EPCs). Methods Mononuclear cells from rat bone and spleen were isolated by density centrifugation and cultured in vitro. Successful rate for culturing EPCs was observed in a different condition, including CO2 concentration of culture box(5%,5. 5%),Ph in DMEM-L medium (Ph 7. 2, Ph 7. 4) ,the serum concentrations of 10% ,20% in medium, one month and two month old rat and bone or spleen derived or bone+ spleen derived-EPCs respectively. Results Our results showed that successful rate for culturing EPCs in 5. 5% CO2 concentration of culture box group was higher than 5% CO2 concentration of culture box group(P<0. 05). In addition,the higher successful rate for culturing EPCs was also observed in Ph 7. 2 of DMEM-L medium(P<0. 05). At the same time, successful rate of one month old group was hither than two month old group(P<0. 05). Interestingly,successful rate of different serum concentration group and different derived-EPCs group were not statistically signifcant(P>0. 05). Conclusion It is an optimal culture condition of 5. 5% CO2 concentration of culture box and Ph 7. 2 of DMEM-L medium for EPCs culturing,and the younger rat, the more successful rate of EPCs culturing.%目的 探讨培养大鼠内皮祖细胞的最佳适宜条件.方法 通过密度梯度离心法分离培养来源于大鼠骨髓及脾脏的单核细胞,观察在不同条件下[pH值、二氧化碳(CO2)浓度、血清浓度、取材大鼠月龄、内皮祖细胞来源]培养内皮祖细胞的成功率.结果 pH 7.两组内皮祖细胞培养的成功率明显高于pH 7.4组(P＜0.05),5.5% CO2浓度组内皮祖细胞培养的成功率明显高于5.0% CO2浓度组(P＜0.05),1月龄组内皮祖细胞培养的成功率明显高于2月龄组(P＜0.05),血清浓度及内皮组细胞来源对细胞培养的成功率无明显影响(P＞0.05).结论 大鼠内皮祖细胞适宜在pH 7.2 的DMEM-L培养液及培养箱CO2浓度为5.5%的环境下生长,且年幼的大鼠取材培养的成功率高.