热灭活补体活性的血清对间充质干细胞和免疫细胞培养的优化效应

Optimal Effects of Heat-Inactivated Complement Serum on Mesenchymal Stem Cell and Immune Cell Cultures

该研究目的在于评估胎牛血清热灭活对细胞培养的影响。对经56°C、30 min热灭活前后的血清配制培养基培养的人脐带来源间充质干细胞(human umbilical cord mesenchymal stem cells,hUCMSCs)和人外周血来源单个核细胞(peripheral blood mononuclear cells,PBMCs),进行增殖速率、免疫调控蛋白表达水平、T细胞增殖速率的检测,比较血清热灭活对细胞培养的影响。结果显示,热灭活前后的两种血清对hUCMSCs的形态没有影响,但热灭活处理能够提高hUCMSCs增殖速率以及响应炎症因子的能力从而使其表达更多的免疫调控蛋白IDO(indoleamine 2,3-dioxygenase)和COX-2(cyclooxygenase 2);此外,热灭活的血清能够显著提高T细胞的体外增殖速率。补体系统活性的标志蛋白C3a在热灭活血清中显著下调。向含有未热灭活血清的培养基中加入补体活性抑制剂compstatin,同样能够提高hUCMSCs的增殖速率和免疫抑制能力。综合以上结果,该文研究表明,56°C、30 min热灭活血清处理能够通过热灭活补体系统提高干细胞的增殖能力、免疫调控效应以及T细胞的增殖速率,建议在科研实验中使用热灭活的血清以优化细胞培养程序。

This study was aimed to evaluate the effect of heat-inactivated fetal bovine serum on cell culture.Umbilical cord derived mesenchymal stem cells and human peripheral mononuclear cells were cultured in the medium with uninactivated and heat-inactivated serum in 56°C for 30 min.There were significant differences between uninactivated and heat-inactivated serum in regard to the rate of proliferation of mesenchymal stem cell,levels of immunoregulatory proteins and rate of proliferation of T cells.The results showed that there was no difference in the morphology of human umbilical cord-derived mesenchymal stem cells cultured with heat-treated or untreated serum,but the proliferation rate and expression levels of immune regulatory proteins IDO and COX-2 were increased by cultured with heat-inactivated serum.In addition,heat-inactivated serum could significantly increase the proliferation rate of T cells.Additionally,complement C3a,which reflected the activity of complement system,was downregulated after heat inactivation.However,the proliferation of hUCMSCs cultured in medium contained uninactivated serum was also increased through inhibiting the activity of complement using compstatin.Besides that,compstatin treatment also upregulated the IDO and COX-2 expression levels cultured in uninactivated serum contained medium.In conclusion,heat-inactivated serum at 56°C for 30 min could enhance the immune regulation ability of stem cells and increase the proliferation rate of T cells.Therefore,heat-inactivated serum was recommended in scientific research experiments.