大鼠骨髓间充质干细胞条件培养液对心脏成纤维细胞胶原合成的影响

目的体外模拟机体缺血环境,研究骨髓间充质干细胞(MSCs)旁分泌对心脏成纤维细胞胶原合成的影响,为MSCs移植机制提供实验依据。方法分离培养SD大鼠的MSCs,换以无血清培养液同时缺氧处理不同时间,然后收集MSCs的条件培养液,以此条件培养液作为刺激因子孵育 SD大鼠的心脏成纤维细胞,用MTT和3H-脯氨酸掺入观察心脏成纤维细胞的增殖及胶原合成。结果用MSCs条件培养液培养心脏成纤维细胞,3H-脯氨酸掺入明显高于对照组,缺氧6 h组的3H-脯氨酸掺入高于对照组约100％,提示MSCs条件培养液刺激心脏成纤维细胞胶原合成增加;MTT结果无显著差异,提示:MSCs条件培养液没有影响心脏成纤维细胞的增殖。结论大鼠骨髓间充质干细胞的缺氧和无血清条件培养液能够通过旁分泌刺激心脏成纤维细胞自身合成胶原能力的增强,提示移植到缺血心肌缺血区的骨髓间充质干细胞可能通过旁分泌作用影响心脏成纤维细胞的胶原合成,从而参与损伤心肌的修复。

Effects of Conditioned Media of Rat Mesenchymal Stem Cells on Proliferation and Collagen Synthesis of Cultured Cardiac Fibroblasts in Neonate Rat

Objective To further examine the paracrine role of MSCs in cell transplantation,MSCs were exposed to the condition that mimicked ischemia and the effects of MSCs-conditioned media ( MSCCM ) on proliferation and collagen synthesis of cardiac fibroblasts ( CFBs) were studied. Methods MSCs from SD rats were cultured with serum-free media under hypoxic conditions for 3 ,6,9,24 hours and then the conditioned media were collected separately. Cultured CFBs of neonate rat were stimulated with the above MSCCM. Cell proliferation was measured by MTT assay.3H-proline incorporation was used to assay the collagen synthesis of CFBs. Result The incorporation of 3H-proline was significantly higher in MSCCM stimulated group than in the control group, indicating that MSCCM promoted collagen synthesis of CFBs. MTT assay indicated that MSCCM did not influence the proliferation of CFBs. Conclusion MSCCM enhanced the collagen synthesis of CFBs but had no effect on the proliferation of CFBs. These results suggested that the paracrine of MSCs may play an important role in MSCs transplantation and may participate in repairing injured myocardium.