【摘要】：目的:探討鹿茸多肽(PAP)聯合膠質細胞源性神經營養(yǎng)因子(GDNF)基因修飾的雪旺細胞對人骨髓間質干細胞(BMSCs)體外增殖的影響。方法:按常規(guī)方法抽取10mL健康志愿者骨髓,接種于培養(yǎng)瓶中進行培養(yǎng),原代培養(yǎng)細胞完全融合前進行傳代,傳至第3代時收獲BMSCs,調整細胞濃度為5×106 mL-1備用。加入4μL GDNF基因修飾的雪旺細胞為GDNF組,加入4μL(10mg·L-1)PAP聯合GDNF基因修飾的雪旺細胞作為聯合組,對照組未加入任何藥物僅加入等量的培養(yǎng)基。采用酶聯免疫檢測法檢測各組細胞細胞增殖活力,ELISA法檢測各組BMSCs中增殖細胞核抗原(PCNA)水平,AnnexinⅤ-FIFC/PI細胞凋亡檢測試劑盒檢測細胞凋亡率。結果:原代培養(yǎng)48h后大部分細胞貼壁,形態(tài)轉變?yōu)槎嘟切?少數呈梭形。傳代細胞幾乎呈梭形,為BMSCs,且均為非造血干細胞。與對照組比較,GDNF組和聯合組細胞增殖活力和BMSCs中PCNA水平升高(P0.05),細胞凋亡率降低(P0.05);與GDNF組比較,聯合組細胞增殖活力和BMSCs中PCNA水平升高(P0.05),細胞凋亡率降低(P0.05)。結論:PAP聯合GDNF基因修飾的雪旺細胞對人BMSCs體外增殖具有明顯的促進作用。
[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.
【作者單位】： 長春中醫(yī)藥大學臨床醫(yī)學院;
【基金】：吉林省科技廳科技發(fā)展計劃項目資助課題(20150520049JH)
【分類號】：R318.08;R68

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