异源性心肌细胞裂解液诱导骨髓干细胞免疫源性的实验研究

Porcine MSC-derived CLC induced by rabbit's cardiomyocytes extract did not induce immune reaction in vivo

目的将家兔心肌细胞裂解液体外诱导小型猪骨髓间充质干细胞(MSC)所得心肌样细胞回输入原动物,观察其是否激活机体免疫反应。方法将小型猪 MSC 体外经家兔心肌细胞裂解液诱导所得的心肌样细胞与单纯培养基培养的 MSC 分别回输入原小型猪体内,于不同时间点,采用流式细胞计数法检测外周血 CD4^+、CD8^+T 淋巴细胞数目变化,ELISA 法检测体外血清白介素(IL)-2、IL-4浓度变化,采用 Cell Counting Kit-8试剂盒检测细胞回输后小型猪脾淋巴细胞细胞毒性 T淋巴细胞(CTL)杀伤活性。结果两组细胞回输入原小型猪体内后血 CD4^+、CD8^+T 淋巴细胞数目,血清 IL-2、IL-4浓度,在各时间点间差异均无统计学意义(P均>0.05),各组动物脾淋巴细胞 CTL 杀伤活性差异无统计学意义。结论经家兔心肌细胞裂解液诱导所得心肌样细胞回输入原小型猪体内,不激活机体免疫反应,提示采用此方法制备的心肌样细胞可能有较好的应用前景。

Objective To compare the in vivo immune reaction of transplanting porcine MSC- derived CLC with rabbit cardiomyocytes extracts induced differentiation or in vitro cultured porcine MSC. Methods After injecting the MSC-derived CLC or MSC to the original porcines, the number of CD4^＋ , CD8 ^＋T cells were determined by flow cytometry. The serum concentrations of IL-2, IL-4 were measured by ELISA, and the porcine spleen lymphocyte CTL cytotoxicity was determined by Cell Counting Kit-8 Assay. Results The numbers of CD4 ^＋ and CD8 ^＋T cells, the serum concentrations of IL-2, IL-4 and spleen lymphocyte CTL cytotoxicity were all similar in porcines received MSC-derived CLC induced by rabbit＇s CMs extract or MSC transplantation. Conclusion The porcine MSC-derived CLC induced by rabbit＇s CMs extract did not induce extra immune reaction when injected back to the original porcine.