ABSTRACT
Background: Many autocrine and paracrine elements that are produced within follicular niche have been the focus of much in vitro maturation [IVM] research. The present study was carried out to compare retinoic acid [RA] and basic fibroblast growth factor [bFGF] efficacy on IVM of mouse oocytes, and their further dual consumption to reach an optimal protocol
Materials and Methods: In this experimental study, germinal vesicle [GV] oocytes obtained from two-months-old NMRI mice were randomly divided into control, sham and three experimental groups. The basic culture medium was alpha-MEM supplemented with 10% fetal bovine serum [FBS], 50 mg/l streptomycin, 60 mg/l penicillin and 10 ng/ ml epidermal growth factors. Each of the experimental groups received one of the following treatments: RA [2 microM], bFGF [20 ng/ml] or combination of RA and bFGF with the indicated concentrations. After 24 hours, capacitated spermatozoa were added to in vitro matured oocytes. Five hours later, the oocytes were cultured in fresh droplets of M2 medium for 24 hours and assessed for cleavage to the two-cells stage
Results: As compared with the control group, the rate of maturation was significantly increased in the RA [P<0.001] and bFGF+RA [P<0.02] groups with 58 +/- 10 and 57 +/- 3.46, respectively. The rate of maturation was significant in the RA [P<0.02] and bFGF+RA [P<0.03] groups, in comparison with the bFGF group. The bFGF+RA group had higher rate [83 +/- 1.52] of two-cells development, than control [33 +/- 1, P<0.001]
Conclusion: Our findings showed beneficial effects of 2 micro M RA and 20 ng/ml bFGF combination on mouse oocyte IVM
ABSTRACT
Objective: Bone marrow has recently been recognized as a novel source of stem cells for the treatment of wide range of diseases. A number of studies on murine bone mar-row have shown a homogenous population of rare stage-specific embryonic antigen 1 [SSEA-1] positive cells that express markers of pluripotent stem cells. This study focuses on SSEA-1 positive cells isolated from murine bone marrow in an attempt to differentiate them into insulin-secreting cells [ISCs] in order to investigate their differentiation potential for future use in cell therapy
Materials and Methods: This study is an experimental research. Mouse SSEA-1 positive cells were isolated by Magnetic-activated cell sorting [MACS] followed by characterization with flow cytometry. Induced SSEA-1 positive cells were differentiated into ISCs with specific differentiation media. In order to evaluate differentiation quality and analysis, dithizone [DTZ] staining was use, followed by reverse transcription polymerase chain reaction [RT-PCR], immunocytochemistry and insulin secretion assay. Statistical results were analyzed by one-way ANOVA
Results: The results achieved in this study reveal that mouse bone marrow contains a population of SSEA-1 positive cells that expresses pluripotent stem cells markers such as SSEA-1, octamer-binding transcription factor 4 [OCT-4] detected by immunocytochemistry and C-X-C chemokine receptor type 4 [CXCR4] and stem cell antigen-1 [SCA-1] detected by flow cytometric analysis. SSEA-1 positive cells can differentiate into ISCs cell clusters as evidenced by their DTZ positive staining and expression of genes such as Pdx1 [pancreatic transcription factors], Ngn3 [endocrine progenitor marker], Insulin1 and Insulin2 [pancreaticbeta-cell markers]. Additionally, our results demonstrate expression of PDX1 and GLUT2 protein and insulin secretion in response to a glucose challenge in the differentiated cells
Conclusion: Our study clearly demonstrates the potential of SSEA-1 positive cells to differentiate into insulin secreting cells in defined culture conditions for clinical applications
ABSTRACT
Diabetes Mellitus [DM], simply known as diabetes, refers to a group of metabolic diseases in which there are high blood sugar levels over a prolonged period. In this study, the feasibility and safety of intravenous transplantation of Very Small Embryonic Like stem cells [VSELs] were investigated for diabetes repair, and finally the migration and distribution of these cells in hosts were observed. Mouse bone marrow VSELs were isolated by Fluorescent Activating Cell Sorting [FACS] method by using fluorescent antibodies against CD45, CXCR4 and Sca1 markers. Sorted cells were analyzed for expression of oct4 and SSEA1 markers with immunocytochemistry staining method. To determine multilineage differentiation, sorted cells were differentiated to Schwann, osteocyte and beta cells. Ten days after the establishment of a mouse model of pancreas necrosis, Dil-labeled VSELs were injected into these mice via tail vein. Pancreases were harvested 4 weeks after transplantation and the sections of these tissues were observed under fluorescent microscope. It was proved that CD45-, CXCR4+, and Sca1+ sorted cells express oct4 and SSEA1. Our results revealed that intravenously implanted VSELs could migrate into the pancreas of hosts and survive in the diabetic pancreas. In treated groups, blood glucose decreased significantly for at least two month and the weights of mice increased gradually. This study provides a strategy for using VSELs for curing diabetes and other regenerative diseases, and the strategy is considered an alternative for other stem cell types
Subject(s)
Animals, Laboratory , Diabetes Mellitus/therapy , Mice , Leukocyte Common Antigens , Receptors, CXCR4 , Pancreas , Injections, IntravenousABSTRACT
Numerous in vitro reports suggest that Low Level Laser Therapy [LLLT] affects cellular processes by biostimulation, however most of them emphasize on using visible light lasers which have low penetration. The aim of this study was to determine the effect of infrared laser light [which is more useful in clinic because of its higher penetration] on secretion of Fibroblast Growth Factor [FGF], Platelet Derived Growth Factor [PDGF] and Vascular Endothelial Growth Factor [VEGF], as important growth factors in wound healing. Fibroblasts were extracted from the skin of 7 diabetic and 7 nondiabetic mice and cultured. Cell cultures of experimental group were irradiated with single dose of LLLT [energy density of 1 J/ cm[2]] using an 810 nm continuous wave laser and the control group was not irradiated. Secretion of growth factors by skin fibroblasts were quantified through real time polymerase chain reaction. Diabetic irradiated group showed significant increase in FGF [p=0.017] expression, although PDGF increased and VEGF decreased in both diabetic and nondiabetic irradiated groups, but these variations were not statistically significant. These results suggest that LLLT may play an important role in wound healing by stimulating the fibroblasts