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A Preliminary Study of the Effects of PGE1 on Cell Proliferation of Human Adipose Derived Stem Cells

Author: YangHuanNa
Tutor: CuiZhengJun
School: Zhengzhou University
Course: Surgery
Keywords: PGE1 ADSCs cell separation cell proliferation
CLC: R329
Type: Master's thesis
Year: 2011
Downloads: 25
Quote: 0
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Abstract


Background and ObjectiveSubcutaneous soft tissue filling plastic surgery has been a common treatment, the surface of the skin due to soft tissue fullness, depression, abnormal contraction present, the common or autologous free fat filling all kinds of artificial materials. Rejection due to the existence of artificial materials, such as trauma and safety problems which are difficult to meet clinical needs, and free autogenous fat graft survival rate of only 40% to 50%, which makes its clinical application is greatly limited. Absorption of fat grafts is one of the reasons is the mature fat cells to hypoxia, damage tolerance is low, and not the proliferation of updates. The adipose derived stem cells (ADSCs) is a strong potential for proliferation and differentiation, greater tolerance to hypoxia, and autologous stem cell transplantation for the town to avoid rejection. More secure.Just like bone marrow mesenchymal stem cells(BM-MSCs), aidpose derived stemcells(ADSCs)have the capacity of multi-differentiation. However,ADSCs can be isolatedeasily with abundant rcsoBrces and little injury to patients. Thus more and more researchersbecame interested in the study of ADSCs. One of the important applications of ADSCs is Tissue Engineering. It’s very clear that ADSCs are the most suitable seed cells for the construction of engineering adipose tissues. Thus, it is meaningful to study the proliferation and differentiation of ADSCs.prostaglandinEl (PGE1) as a peotect factor derived from vascular endothelial cells, It is an effective vasodilating agent. For a long time, PGE1 is apply to the treatmem of pulmonary artery hypenension. With me development of clinical application and research, it is gradually realized that PGE1 can inhibit platelet aggregation and mrombogenesis, relax the smooth muscle of bronchus, prevent thrombogenesis, improve ischemic myocardium, inhibit the degranulation of macrophage and blood corpuscle, improvered blood cell defomation and has positive inotropic effect. Which is generally used to coronary heart disease, heart failure, hyperlipemia, diabetic neuropathy and chronic renal insuffciency.Studies have found that misoprostol (PGE1 analogue) on human preadipocyte proliferation and differentiation has a strong stimulating effect, while the effect of PGE1 on the proliferation of ADSCs has not been reported, of prostaglandins on the proliferation of stem cells and Differentiation effect on the quest to improve survival rate of transplanted adipose derived stem cells clinically practical significance. Therefore the design of the experiment, preliminary exploration PGE1 on cell proliferation effect of ADSCs.MethodsThe mesenchymal stem cells in adipose tissue were extracted from abdomen wall of 4 patients, donors undergoing the lipesuction in clinic, then these cells were induced to differentiate to the adipogenic cells in vitro. Cell morphology was observed under the microscope, Flow cytometric analysis of the cell surface antigens. When the adipose derived stem cells in an excessive number of one-time training, short term and will not be fully used, it can be part of the cryopreserved for future use of the experiment and then recovery. The cultured ADSCs were treated with five different concentration of PGE1, which were 3μmol/L,6μmol/L,9μmol/L,12μmol/L, and 0μmol/L (control group). After 3 days, observe the cell morphology and proliferation. ResultsADSCs into the culture dish 48 hours after planting a large number of adherent spindle cells, showing fibroblast-like growth, rich in cytoplasm and nucleolus, cell morphology varies with short processes, were small polygonal or short spindle nuclei Center.5 days, cells reached the logarithmic growth phase, growth of spindle cells fused together, closely packed a certain direction, or parallel-like arrangement was swirling.9 days after cell confluence culture bottles, each colony was fused bundle or whirlpool order.13 days later, a typical fibroblast-like growth. ADSCs flow cytometry showed that the relative expression of specific molecules CD44-, CD105, CD34, CD45, CDHLA-DR. By experimental observation and testing to show that the cells harvested in this experiment indeed form a complete, fully functioning stem cells, There was no obervious different among the experimental groups. And the cell number of each experimental group was obervious elevated, comparing with control group, The difference was statistically significant (p<0.05), Compared to the experimental group, some differences were statistically significant (the experimental group compared with group p=0.004).Conclusions1. The enzyme digestion experiments using cultured cells isolated by histology and identification of surface markers indeed adipose-derived stem cells;2. Adipose-derived stem cells were treated with different concentrations of PGE1, the number of cells showed an increasing trend, according to statistics, there was significant difference between the groups, the initial description of the PGE1 can proliferation of adipose-derived stem cells.3. Application of statistical software to compare the group discovered that some of the differences between the experimental group was statistically significant, indicating that PGE1 concentrations reach a certain height, cell number and degree of value-added PGE1 in a dose dependent manner.

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