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人神经生长因子β在兔骨髓间充质干细胞中的重组表达。

Recombinant expression of human nerve growth factor beta in rabbit bone marrow mesenchymal stem cells.

机构信息

Department of Internal Neurology, Second Affiliated Hospital, Zhengzhou University, No. 2 JingBa Road, Zhengzhou, Henan, 450014, People's Republic of China.

出版信息

Mol Biol Rep. 2010 Dec;37(8):4083-90. doi: 10.1007/s11033-010-0068-4. Epub 2010 Mar 17.

Abstract

Nerve growth factor (NGF) is required for the differentiation and maintenance of sympathetic and sensory neurons. In the present study, the recombinant expression of human nerve growth factor beta (hNGF-β) gene in rabbit bone marrow mesenchymal stem cells (rMSCs) was undertaken. Recombinant vector containing hNGF-β was constructed and transferred into rMSCs, the expressions of the exogenous in rMSCs were determined by reverse transcriptase PCR (RT-PCR), ELISA and Western blot, whereas the biological activity of recombinant hNGF-β was confirmed using PC12 cells and cultures of dorsal root ganglion neurons from chicken embryos. The results showed that the hNGF-β gene expressed successfully in the rMSCs, a polypeptide with a molecular weight of 13.2 kDa was detected. The maximal expression level of recombinant hNGF-β in rMSCs reached 126.8012 pg/10(6) cells, the mean concentration was 96.4473 pg/10(6) cells. The recombinant hNGF-β in the rMSCs showed full biological activity when compared to commercial recombinant hNGF-β.

摘要

神经生长因子(NGF)是交感神经和感觉神经元分化和维持所必需的。在本研究中,我们进行了人神经生长因子β(hNGF-β)基因在兔骨髓间充质干细胞(rMSCs)中的重组表达。构建了含有 hNGF-β 的重组载体并转染 rMSCs,通过逆转录 PCR(RT-PCR)、ELISA 和 Western blot 检测外源性基因在 rMSCs 中的表达,同时使用 PC12 细胞和鸡胚背根神经节神经元培养物来验证重组 hNGF-β 的生物学活性。结果表明,hNGF-β 基因在 rMSCs 中成功表达,检测到分子量为 13.2 kDa 的多肽。rMSCs 中重组 hNGF-β 的最大表达水平达到 126.8012 pg/10(6) 个细胞,平均浓度为 96.4473 pg/10(6) 个细胞。与商业重组 hNGF-β 相比,rMSCs 中的重组 hNGF-β 具有完全的生物学活性。

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