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find Author "SHAN Genfa" 3 results
  • Protect Effect of Basic Myocardial Construction in Remnant Myocardium by Umbilical Cord Blood Mesenchymal Stem Cells implanted into Infarcted Myocardium

    Objective To investigate the influence of infarcted myocardial construction by umbilical cord blood mesenehymal stem cells(MSC) with induced to myo-derived stem cells and implanted into infarcted myocardium. Methods Thirty-six adult mongrel canines were randomly divided into MSC transplant group and control group (18 each group). Transplant group: the umbilical cord blood MSC differented to myo-derived stem cells induced by 5-azacytidine(5-aza) were implanted into the acute myocardial infarct site via the ligated left anterior descending (LAD) artery. Control group: administer the same volume of IMDM culture medium containing 0. 02% 4,6- diamidino-2-phenylindone (DAPI). At 2, 4, 8 weeks after implantation, the change of basic myocardial construction and the distribution of desmin were observed by using Nagar-Olsen staining and immunohistology respectively. Results With less fusing, the arrangement of gelatine fibers and elastic fibers were in order in transplant group,and they were partly fused in control group by Nagar-Olsen staining. The expression of desmin of infarcted myocardial cell in transplant group was much higher than those in control group (P〈0. 01). No significant difference was detected in the expression of desmin in normal site of both groups (P〉0. 05). Conclusion There is an protective effect on the basic myocardial construction in infarcted myocardium after the umbilical cord blood MSC was differented to myo-derived stem cell by induced with 5-aza in vitro and implanted into the acute myocardial infarction.

    Release date:2016-08-30 06:18 Export PDF Favorites Scan
  • Isolation, Induction and Culture of Human Endothelial Progenitor Cells in Bone Marrow and Amplification in Vitro

    Abstract: Objective To investigate and improve the method of isolation, induction and culture, amplification in vitro of human endothelial progenitor cells (EPCs) in human bone marrow, thus establish a foundation for EPCs to participate in basic research and clinical application. Methods WHuman bone marrow mononuclear cells (hBMMNCs) were isolated by density gradient centrifugation and cultured in the 6plateds coating human fibronections(HFN group), coating gelatinum (coating gelatinum group) and coating nothing (coating nothing group) respectively. After culturing for 4-7d endothelial cell basal medium-2(EBM-2) cell colonyforming units (CFUs) appeared, then select EPCsliked CFUs for cultivation which was named the pick method, CD34+ KDR+ and CD133+ KDR+ double positive cells were detected in flow cytometry, and CD133, CD34, CD31, vWF and KDR expression were detected with cell immunochemical test. Results hBMMNCs were isolated from human bone marrow more effectively with OptiprepTM cell separating medium, and induced and obtained more EPCs, and cultured and amplificated in vitro. Flow cytometer showed CD133+ KDR+ double positive cells reaching up to 70.4%±5.4%, CD34+ KDR+ double positive cells reaching up to 69.1%±8.7%. EPCs grew vigorously in coating HFN group and coating gelatinum group, both HFN and gelatinum promote EPCs adherence and growth, but there were no statistically difference in two groups (Pgt;0.05).Surface mark of adherent cells cultured 7d such as CD133, CD31, vWF and KDR showed positive, and cells cultured 14d such as CD34 showed positive. Conclusion The method of picking CFUs can obtain more EPCs from human bone marrow with success and can amplificate EPCs in vitro, thus introducing another simple and effective method to purify EPCs, further widening range and increasing method to purify EPCs.

    Release date:2016-08-30 06:08 Export PDF Favorites Scan
  • The Cellcell Junction after Implantation of the Myocardiumlike Cell Derived from the Canine Umbilical Cord Blood

    Abstract: Objective To study the integration of transplanted cells and host cells by detection of the cellcell junction after transplantation of the myocardiumlike cell derived from the canine umbilical cord blood. Methods The mesenchymal stem cell(MSCs) was transfected by Laz-Z after harvest, culture, induced by 5-azacytidine(5-aza). Thirty-six adult hybrid dogs were randomly divided into cell transplantation group and control group. The canine of myocardium infarction was established. 107 MSCs were transplanted into dogs with acute myocardium infarction by coronary artery infusion and local injection in cell transplantation group and physiologic saline was used in the control group. The specimens were harvested and detected by immunofluorescence for 2, 4 and 8 weeks respectively. Results The umbilical cord blood MSCs were fusiform or spindleshaped. They presented clonal and knittinglike growth.The MSCs could differentiate into myocardium-like cell by the induction of 5-aza and express α-actin, desmin, connexin43.The transplanted cells could survive more than 8 weeks after transplantation. Cadherin and connexin 43 were found in the position of cellcell junction of transplanted cells group and between transplanted cells and host cells. Cadherin and connexin 43 were found in the hose cells of the control group. Conclusion The umbilical cord blood MSCs is able to differentiate into myocardiumlike cell in vitro and form cellcell junction in vivo to communicate with surrounding cells.

    Release date:2016-08-30 06:08 Export PDF Favorites Scan
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