Human preproenkephalin gene-modified bone marrow mesenchymal stem cell line:its biological characteristics

Li Xin-mao

doi:10.3969/j.issn.2095-4344.2016.19.001

Abstract：BACKGROUND:Previous studies have found that the transplantation of modified cel lines exhibit analgesic effect. But little is reported on the biological characteristics of human preproenkephalin gene-modified bone marrow mesenchymal stem cel lines. OBJECTIVE:To observe the biological characteristics of human preproenkephalin gene-modified bone marrow mesenchymal stem cel s. METHODS:Bone marrow mesenchymal stem cel s were isolated and cultured to establish human preproenkephalin gene-modified bone marrow mesenchymal stem cel lines. RESULTS AND CONCLUSION:After freezing-thawing, passage 4 human bone marrow mesenchymal stem cel s, human bone marrow mesenchymal stem cel s-pBABE and human bone marrow mesenchymal stem cel s-human preproenkephalin exhibited no significant changes in the cel viability (P>0.05), as wel as in the fat cel proportion after adipogenic induction (P>0.05). Moreover, red calcium deposition was presented in al these cel s by alizarin red staining after osteogenic induction. Flow cytometry results showed that passage 4 human bone marrow mesenchymal stem cel s, human bone marrow mesenchymal stem cel s-pBABE and human bone marrow mesenchymal stem cel s-human preproenkephalin could express CD29 and CD44, but not express CD34 and CD45. Human preproenkephalin genes were highly expressed in human bone marrow mesenchymal stem cel s-pBABE, but lowly expressed in passage 4 human bone marrow mesenchymal stem cel s. Additional y, recombinant plasmid pBABE-preproenkephalin-modified human bone marrow mesenchymal stem cel s could express enkephalin protein. In conclusion, human preproenkephalin gene-modified human bone marrow mesenchymal stem cel lines can maintain the pluripotent differentiation or proliferation capacity of bone marrow mesenchymal stem cel s, and secrete enkephalin protein.