Mesenchymal Stem Cells with Calreticulin Gene Modulate Cell Adhesiveness Through an Integrin-mediated Mechanism
의약학 > 의공학
( Yang Soo Jang ) , ( Byoung Hyun Min ) , ( Sun Ju Lee ) , ( Nam Sik Chung ) , ( Hye Jung Kim ) , ( Ki Chul Hwang ) , ( So Yeon Lim ) , ( Woo Chul Chang ) , ( Byeong Wook Song ) , ( Min Ji Cha ) , ( Sae Yun Kwon ) , ( Seong Min Han ) , ( Jong Youn Kim )
조직공학과 재생의학 2006년, 제3권 제3호, 327~335페이지(총9페이지)
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    Transplantation of adult bone marrow-derived mesenchymal stem cell(MSC) has the potential to treat the cardiac repair following myocardial injury. However, poor cell viability due to anoikis, cell death induced by the loss of cell/matrix interaction, after transplantation has limited the regenerative capacity of these cells in vivo. It is known that adhesion is a key factor for the differentiation of MSCs. In this study, we genetically modified the rat mesenchymal stem cells with calreticulin(CRT) ex vivo to enhance the adhesiveness to the cardiac matrix and survival in the last. We used MSCs with fibroblast-like morphology after 2 weeks culture, which was expressed CD71 over 95%. CRT-transfected MSCs expressed over 8-fold higher of CRT than non-transfected control MSCs. Quantitative adhesion experiments demonstrated that CRT transfection increased adhesion of MSCs to fibronectin. Similarly, migration of CRT-transfected MSCs(CRT-MSCs) was increased at the same manner. On the other hand, down-regulation of CRT by transfection of CRT-siRNA significantly decreases adhesion and spreading of MSCs on fibronectin. Likewise, interfering with the adhesive function of CRT markedly reduces migration of MSCs on fibronectin. CRT-MSCs on fibronectin displayed a significant increase in phosphorylation of focal adhesion related genes, FAK and Src, compared to MSCs only. Phosphorylation of PI3K was also increased in CRT transfected MSCs. 1 week after the induction of rat myocardial infarction using LAD ligation, MSCs labeled with 4`,6-diamino- 2-phenylindole(DAPI) were implanted. After 1 week, survival of CRT-MSCs was 30% higher than that of control MSCs and in the former, migration of implanted cells was enhanced. Our studies indicate that CRT-MSCs enhance cell adhesiveness into a matrix as well as spreading and migration and also enhance the assembly of focal adhesion complexes in vitro. These data may provide genetic modification of MSCs with a therapeutic gene before transplantation as novel therapeutic rationales for the treatment of myocardial infarction and end-stage cardiac failure.
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