Recombinant expression of human nerve growth factor beta in rabbit bone marrow mesenchymal stem cells.
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.
Fan
Bo-Sheng
BS
Department of Internal Neurology, Second Affiliated Hospital, Zhengzhou University, No. 2 JingBa Road, Zhengzhou, Henan, 450014, People's Republic of China.
Lou
Ji-Yu
JY
D000818:Animals; D001483:Base Sequence; D001681:Biological Assay; D001854:Bone Marrow Cells / Q000166:cytology*; D002469:Cell Separation; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000187:drug effects; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008969:Molecular Sequence Data; D020932:Nerve Growth Factor / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D016501:Neurites / Q000187:drug effects; D016716:PC12 Cells; D010957:Plasmids / Q000235:genetics; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection; D019076:Transgenes
D011994:Recombinant Proteins; D020932:Nerve Growth Factor