[Differentiation potential of EGFP gene-modified mesenchymal stem cells transfected by lentiviral vector].
This study was aimed to investigate the differentiation potential after EGFP gene-modified mesenchymal stem cells (MSCs) transfected by lentiviral vector (LV). After isolated, cultured and identified, MSCs were transfected with the lentiviral vector carrying EGFP gene in vitro. The transfection efficiency was measured by observing the expression of green fluorescence protein. At last, the transfected MSCs were induced into adipocytes in adipogenesis supplement medium, the induction level was detected by Sudan fat stain. The results indicated that after transfection for 72 hours, weak fluorescence in MSCs was observed under fluorescence microscope. After 21 days, many lipid droplets with high refractivity occurred in cytoplasm of transfected MSCs, and showed orange in Sudan black stain. There were no significantly differences between transfected and non-transfected cells (p > 0.05). It is concluded that MSCs were successfully transfected by LV carrying EGFP gene. The transfected MSCs maintain multiple differentiation and proliferation potential. In the adipogenesis supplement medium, transfected MSCs also can be induced to differentiate into adipocytes. MSCs can act as target cells for gene therapy.
Chen
Hui-Ling
HL
Department of Hematology, The Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China.
Bai
Hai
H
D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D014162:Transfection
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins