ABSTRACT
BACKGROUND:Islet and islet cel transplantation for the treatment of diabetes has achieved effect, but the research is limited dut to the shortage of islet and immune rejection. OBJECTIVE:To observe the effect of transplantation of islet-like cells that in vitro differentiated from bone marrow mesenchymal stem cells on the treatment of diabetes in rats. METHODS:The rat bone marrow mesenchymal stem cells were induced with basic fibroblast growth factors and hepatocyte growth factors, and then received immunocytochemistry staining to detect the induction. The Sprague Dawley rats received intraperitoneal injection of streptozotocin to establish the diabetes models. After modeling, the rats were randomly divided into control group and experimental group (transplanted with induced islet-like cells). The experimental group was transplanted with the induced islet-like cells through renal capsule, and the control group was transplanted with normal saline in the same dose. The blood glucose and body mass of the diabetes rats were observed after transplantation. RESULTS AND CONCLUSION:The bone marrow mesenchymal stem cells could differentiate into islet-like cells after in vitro induced with basic fibroblast growth factors and hepatocyte growth factors. There was no significant change in blood glucose of the control group after transplantation (P>0.05), and the blood glucose of the rats in the experimental group was significantly decreased compared with the control group (P<0.05). The bone marrow mesenchymal stem cells can differentiate into islet-like cells after in vitro induced with the induction system containing basic fibroblast growth factors and hepatocyte growth factors, and the islet-like cells have a certain ability of insulin secretion. The transplantation of induced islet-like cells after transplanted into the diabetes rats through renal capsule can decrease the blood glucose level of the rats.
ABSTRACT
In order to set up a base for stem cells to be widely used in clinical medicine, we tried to optimize, in this study, the technique that induces human mesenchymal stem cells (hMSCs) to differentiate into neural stem cells by using cerebrospinal fluid (CSF) from the different groups. After the induction, presence of neural stem cells was confirmed with microscope observation, flow cytometry analysis, immunohistochemistry and fluorescent immunohistochemistry. At the same time, we also compared and analysed the data of the number of stem cells when it totally met the requirements for clinical treatment and the days required. At last, we confirmed that hMSCs could be induced to differentiate into neural stem cells, and that the number of cells totally met the requirements for clinical treatment. But there were some differences both in the number of cells and the days required. Among the groups, the group that marrow mesenchymal stem cells from patients own induced by CSF from healthy volunteers used the shortest time and the quantity of the cells was significantly higher than those of the others.
Subject(s)
Humans , Cell Differentiation , Cerebrospinal Fluid , Chemistry , Culture Media , Chemistry , Flow Cytometry , Immunohistochemistry , Mesenchymal Stem Cells , Cell Biology , Neural Stem Cells , Cell BiologyABSTRACT
Objective To study the effects of anti-oncogene WWOX on cell growth of epithelial ovarian cancer,in order to find a new approach of gene therapy for ovarian cancer.Methods A eukaryotic expression vector containing WWOX was transfected into ovarian cancer cell line HO8910 in vitro (recombinant plasmid group),and positive cell clones were selected and amplified.Expression of WWOX protein was detected by western blot. Untransfected cell(blank contrast group) and transfected empty plasmid cell(empty plasmid group)were served as control groups.In vitro,the biology effect of WWOX on HO8910 cell was analyzed throush the methyl thiazolyl tetrazolium test,transwell chamber cell invasion assay in vitro,agarose clony-formation and flow cytometry.In vivo,the cell of transfection was transplanted intraperitoneally in to BALB/c nude mice.The survival time and growth ability of nude mice were observed.Results (1)Recombinant plasmid group cell could steadily express WWOX protein,while in empty plasmid group and blank control group the expression of WWOX protein were not detected.(2)The growth rate of recombinant plasmid group cell was inhibited.(3)The agnrose clony-formation rate of recombinant plasmid group(19.8%)was significantly lower than that of the empty plasmid group(54.5%)and blank control group(56.0%,P＜0.05).(4)Flow cytometry showed that(72.08±0.39)% of cells was arrested at G0/G1 stage in recombinant plasmid group, while in empty plasmid group and blank control group G0/G1 stage cells were at (41.02±1.08)% and (39.31±0.67)% (P＜0.05). (5) In vitro invasion assay showed that invasion cell number in recombinant plasmid group (89.7±3. 1 ) was not significantly different from that of empty plasmid group(91.2±1.3) and blank control group(91.4±1.3, P ＞0. 05). (6) In vivo test in nude mice showed that WWOX gene could inhibit tumor growth of the HO8910 cells. Conclusions Tumor suppressor gene WWOX could interfere with the cell cycles of ovarian cancer cell and inhibit cell proliferation. As a new valuable tool,it premises to have application in the gene therapy of ovarian cancer.