鹿茸多肽联合GDNF基因修饰的雪旺细胞对人骨髓间质干细胞体外增殖的影响
发布时间：2019-07-08 19:15
【摘要】：目的:探讨鹿茸多肽(PAP)联合胶质细胞源性神经营养因子(GDNF)基因修饰的雪旺细胞对人骨髓间质干细胞(BMSCs)体外增殖的影响。方法:按常规方法抽取10mL健康志愿者骨髓,接种于培养瓶中进行培养,原代培养细胞完全融合前进行传代,传至第3代时收获BMSCs,调整细胞浓度为5×106 mL-1备用。加入4μL GDNF基因修饰的雪旺细胞为GDNF组,加入4μL(10mg·L-1)PAP联合GDNF基因修饰的雪旺细胞作为联合组,对照组未加入任何药物仅加入等量的培养基。采用酶联免疫检测法检测各组细胞细胞增殖活力,ELISA法检测各组BMSCs中增殖细胞核抗原(PCNA)水平,AnnexinⅤ-FIFC/PI细胞凋亡检测试剂盒检测细胞凋亡率。结果:原代培养48h后大部分细胞贴壁,形态转变为多角形,少数呈梭形。传代细胞几乎呈梭形,为BMSCs,且均为非造血干细胞。与对照组比较,GDNF组和联合组细胞增殖活力和BMSCs中PCNA水平升高(P0.05),细胞凋亡率降低(P0.05);与GDNF组比较,联合组细胞增殖活力和BMSCs中PCNA水平升高(P0.05),细胞凋亡率降低(P0.05)。结论:PAP联合GDNF基因修饰的雪旺细胞对人BMSCs体外增殖具有明显的促进作用。
[Abstract]:Aim: to investigate the effect of deer antler polypeptide (PAP) combined with glial cell line-derived neurotrophic factor (GDNF) gene modified Schwann cells on the proliferation of human bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods: the bone marrow of healthy volunteers with 10mL was extracted according to the routine method and inoculated in culture flask for culturing. the primary cultured cells were subcultured before complete fusion, and the adjusted cell concentration of BMSCs, was 5 脳 10 ~ 6 mL-1 when passed to the third generation. The Schwann cells modified by 4 渭 L GDNF gene were added as GDNF group and 4 渭 L (10mg 路L-1) PAP combined with GDNF gene modified Schwann cells as combined group. The control group did not add any drugs to the same amount of culture medium. The cell proliferation activity of each group was detected by enzyme-linked immunosorbent assay (Elisa), the level of proliferating cell nuclear antigen (PCNA) in BMSCs was detected by ELISA assay, and the apoptosis rate of Annexin V-FIFC/PI cells was detected by apoptosis kit. Results: after 48 hours of primary culture, most of the cells adhered to the wall, and the morphology changed into polyhedral shape, and a few of them were fusiform. The passage cells were almost fusiform, BMSCs, and non-hematopoietic stem cells. Compared with the control group, the cell proliferation activity and PCNA level in BMSCs in GDNF group and combination group were increased (P 0.05), and the apoptosis rate was decreased (P 0.05). Compared with GDNF group, the cell proliferation activity and PCNA level in BMSCs in combination group were increased (P 0.05), and the apoptosis rate was decreased (P 0.05). Conclusion: PAP combined with GDNF gene modified Schwann cells can significantly promote the proliferation of human BMSCs in vitro.
【作者单位】： 长春中医药大学临床医学院;
【基金】：吉林省科技厅科技发展计划项目资助课题(20150520049JH)
【分类号】：R318.08;R68
[Abstract]:Aim: to investigate the effect of deer antler polypeptide (PAP) combined with glial cell line-derived neurotrophic factor (GDNF) gene modified Schwann cells on the proliferation of human bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods: the bone marrow of healthy volunteers with 10mL was extracted according to the routine method and inoculated in culture flask for culturing. the primary cultured cells were subcultured before complete fusion, and the adjusted cell concentration of BMSCs, was 5 脳 10 ~ 6 mL-1 when passed to the third generation. The Schwann cells modified by 4 渭 L GDNF gene were added as GDNF group and 4 渭 L (10mg 路L-1) PAP combined with GDNF gene modified Schwann cells as combined group. The control group did not add any drugs to the same amount of culture medium. The cell proliferation activity of each group was detected by enzyme-linked immunosorbent assay (Elisa), the level of proliferating cell nuclear antigen (PCNA) in BMSCs was detected by ELISA assay, and the apoptosis rate of Annexin V-FIFC/PI cells was detected by apoptosis kit. Results: after 48 hours of primary culture, most of the cells adhered to the wall, and the morphology changed into polyhedral shape, and a few of them were fusiform. The passage cells were almost fusiform, BMSCs, and non-hematopoietic stem cells. Compared with the control group, the cell proliferation activity and PCNA level in BMSCs in GDNF group and combination group were increased (P 0.05), and the apoptosis rate was decreased (P 0.05). Compared with GDNF group, the cell proliferation activity and PCNA level in BMSCs in combination group were increased (P 0.05), and the apoptosis rate was decreased (P 0.05). Conclusion: PAP combined with GDNF gene modified Schwann cells can significantly promote the proliferation of human BMSCs in vitro.
【作者单位】： 长春中医药大学临床医学院;
【基金】：吉林省科技厅科技发展计划项目资助课题(20150520049JH)
【分类号】：R318.08;R68
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