| Objective:The aim of this study was to investigate the tissue source of mesenchymal stem cells (MSCs), whether MSCs can be mobilizated from bone marrow and peripheral blood by granulocyte colony-stimulating factor (G-CSF) and to explore the effect of multi-drug factors on the biological characteristics of MSCs and the biocompatibility between MSCs and chitosan.Methods:①MSC derived from adult bone marrow, human fetal bone marrow, and newborn cord and cord blood were isolated by using its adhesiveness. The morphology and the growth curve of MSC were observed, and the surface antigens were analyzed by flow cytometer (FCM). MSC derived from the human uterine cervix cancer tissues were isolated by using its adhesiveness. The morphologic appearance, immunophenotype, growth curve, cell cycle, Chromosome analysis, histochemistry, and differentiation potential of these cells were investigated.②To isolate and culture the MSCs of rat from peripheral blood and marrow being mobilization by granulocyte colony-stimulating factor(G-CSF).③MSCs were exposed to GSH, puerarin and lead acetate. The effects of the three drugs on MSCs were investigated by the determination of cell activity, cell surface markers and cell apoptosis. We also studied the differentiation potency of MSCs though histochemistry and RT-PCR.④MSCs were cultured on the chitosan scaffolds which were prepared by freeze-dryin. We observed cell morphology on the scaffold and detected cell adhesion and cell proliferation.Result:①MSCs were successfully isolated from bone marrow. The reproductive activity of human fetal bone marrow MSCs was obviously more stronger than adult human bone marrow MSCs. Umbilical cord tissue can be successfully isolated MSCs.MSCs couldn't been isolated from newborn cord blood. MSCs were also confirmed to exist in human uterine cervix cancer tissues.②The percentage of CD44+cells increased significantly after mobilization by G-CSF in a dose-dependent manner. MSCs were isolated and cultured unsuccessfully from peripheral blood control group and mobilizated by 20μg/kg G-CSF. The typical colony of MSCs were isolated after being mobilization by 50μg/kg and 80μg/kg G-CSF. Surface marker were to accord with MSCs from bone marrow by FACS assay.③The effect of GSH:0.15mg/ml GSH can promote the growth of MSCs and reduce the apoptosis of MSCs without affecting its own characteristics. The effect of Puerarin:Puerarin can inhibit the proliferation of MSCs in a dose-dependent manner but 0.0012mol/L Puerarin can promote the umbilical cord MSCs differentiation into osteoblasts. The effect of Lead acetate:Lead acetate can inhibit the proliferation of MSCs in a dose-dependent manner, induce cell apoptosis, inhibit osteogenesis differentiation and reduce the ability of secretion of cell factor.④MSCs can adhere and grow on the chitosan scaffolds, and maintain a good cell morphology. Compared with the control group, the adhesion rate and the activity of cell proliferation were not significantly different.Conclusion:①There are MSCs in human fetal bone marrow, umbilical cord, bone marrow, but further study is needed to investigate the existence of MSCs in cord blood. MSCs were also found In cervical cancer tissue. This suggest that MSCs may play an important role in tumor development.②We confirmed that the peripheral blood MSCs from rat could be mobilizated dependent on G-CSF in a given dose.③GSH can promote the growth of MSCs without affecting its own characteristics.this suggest that GSH can be use to improve the rate of in vitro expansion of MSC. Puerarin can inhibit proliferation of MSCs and promote MSCs differentiation into osteoblasts. Lead acetate can influence the activity of proliferation of MSCs, induce cell apoptosis, cause differentiation potential of MSCs decreased. Lead acetate can also reduce the ability of MSCs to secrete cell factors.The damage of MSCs means the damage of hematopoietic microenvironment. This suggest that there may be a certain value in the development of anemia.④There is good biocompatibility between umbilical cord MSCs and chitosan scaffolds.This can be applied to the preparation of composite materials.
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