Construction of a lentivirus vector containing Pax6 and its transfection of human bone marrow mesenchymal stem cells
Liu Jing-wen
Ren Jing
Chen Sheng
Li Bai-jun
Liu Shen-wen
Qin Bo
Abstract：BACKGROUND:Pax6 gene plays an important role in eye development and differentiation, and to study how it regulates the differentiation of human bone marrow mesenchymal stem cel s (BMSCs), gaining the BMSCs stably over-expressing Pax6 is crucial, which is also the basis of stem cel replacement therapy. OBJECTIVE:To construct a lentivirus vector containing Pax6 and detect the expression of Pax6 in transfected human BMSCs. <br> METHODS:Pax6 gene was extracted using PCR. After its connection with lentivirus vector pHIV-EGFP, it was then packaged by 293T cel s. The human BMSCs were transfected with recombinant lentivirus Pax6-EGFP as wel as lentivirus vector pHIV-EGFP, which was considered negative control group. The cel ular morphology was observed by a fluorescence microscope, and the mRNA expression of Pax6 was detected by real-time PCR. <br> RESULTS AND CONCLUSION:The recombinant lentivirus Pax6-EGFP was constructed successful y with a titer of 3×109 pfu/L. After the transfection, both the green fluorescent protein and Pax6 gene were expressed detected using fluorescence microscope and real-time PCR, showing that the method of lentiviral transfection is a safe and effective way to modify BMSCs.
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Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 5338-5344 )