Introduction of hIGF-1 gene into bone marrow stromal cells and its effects on the cell's biological behaviors.
Autologous and gene-modified bone marrow stromal cells (MSCs) have shown a bright future in clinical applications. However, does a gene-modified MSC still maintain its stem cell-like properties? To answer this question, human IGF-1 was introduced into rat MSCs using a recombinant retroviral vector and the effects of the gene manipulation on the cells' behaviors were investigated. The MSCs transfected with hIGF-1 could secrete 6.7-fold higher IGF-1 than the native cells. These MSCs had an elevated baseline activity of ERK signaling, an enhanced proliferation, increased accumulative numbers of cell doublings, and a reduced apoptosis; they showed upregulated expressions of OCT-4, CYP51, and SM22alpha, and a downregulated expression of nestin. This indicates that the overexpressed IGF-1 enhances the MSCs' self-renewal, endodermal and mesodermal differentiation, but weakens their neuronal potential. Although a puromycin selection after hIGF-1 gene transfection could produce a purer transfected MSC population with stronger ability to express functional hIGF-1, it induced premature senescence of the selected cells by activating oncogene Ras, leading to a shortened replicative life span and a weakened multipotency.
Hu
Chengjun
C
Department of Anatomy and Embryology, Wuhan University School of Medicine, Wuhan, PR China.
Wu
Yan
Y
Wan
Yu
Y
Wang
Qiao
Q
Song
Jian
J
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D018014:Gene Transfer Techniques; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics* / Q000502:physiology; D008840:Microfilament Proteins / Q000378:metabolism; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000502:physiology; D009124:Muscle Proteins / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology
D008840:Microfilament Proteins; D009124:Muscle Proteins; D050814:Octamer Transcription Factor-3; C051615:transgelin; D007334:Insulin-Like Growth Factor I; D003577:Cytochrome P-450 Enzyme System; D048049:Extracellular Signal-Regulated MAP Kinases