| Background:In recent years,stem cell therapy is an emerging therapeutic modalities after myocardial ischemia injury repair.A Variety of cell types including embryonic stem cells,induced pluripotent stem cells and adult stem cells candidate as seed cells for the treatment of ischemic heart disease.However,the clinic translation of stem cells therapy faces many restraining factors,such as arrhythmia resulting after transplanting,the oriented differentiation of pluripotent stem cells uncontrollable,tumorigenesis risk and low future fraction surviving of stem cells.In decade years,the trandifferential strategy is expected to solve these problems to some extent.Cell transdifferentiation refers to the lineage switches from one type of differentiated cells to another desired cell type by forcing expression of specific lineage of transcription factors,bypass the pluripotent state.So it has much more efficiency and security.While,introduction of exogenous gene limits the cell-based therapeutic application of transient expressing transcription factors of transdifferential strategies.Because of its rapid biological effects,convenient use and reversible characteristics,small molecules become promising substitute for specific transcription factors in transdifferentiation process.But,the application of small molecules in cell reprogramming is still a new and developing topic,to date the related research of mall molecules strategy were almost isolated,precursor,especially in cardiovascular field.The mechanisms and roadmap of small molecular in the transdifferentiation has yet to be further clarified,more researches need to verify the findings at present,to validate the ability of completely chemical transdifferential to produce one cell types to another only by small molecules.For this concern,the current study is to investigate whether serum deprivation stimulus can activate fibroblasts to endothelial cell transdifferentiation.Methods:Fresh separated heart tissue were used to generation fibroblasts for cell culture form C57 / BL newborn mice.Lymphocyte inhibitory factor(LIF)was added to promote fibroblasts self-renewal,long-term maintain stem property and restrain its trend to terminal differentiation.The third passage of fibroblasts were divided into 5 groups: Control group(DMEM+10%FBS),Serums starvation 24 h(DMEM+0%FBS),Serums starvation 48h(DMEM+0%FBS),Serums starvation 72h(DMEM+0%FBS)and Serums starvation 48 h but without LIF(DMEM+0%FBS).After intervention,their effects on changes of cell lineage specific genes were observed by qPCR.The effects on Fibroblasts angiogenesis was observed by in vitro angiogenesis test.Ac-LDL phagocytic function test was also used to observe Fibroblasts' functional change.Mouse VEGF reagent ELISA kit was used to observe the ability fibroblasts secreting VEGF.Results:The endothelial specific gene CD31 positive cells is 22 times vs control group(P < 0.05)and the express ve-herin positive cells is 50 times vs control group(P < 0.05).Among the three intervention time group,the effect of Serum starvation 48 h is most obvious,the express of ve-herin positive cells is 7.7 times vs Serum starvation 72h(P < 0.05).Contrast to control group,Serum starvation 72 h,it is not obvious change,p>0.05.In addition,Fibroblasts obtained function of endothelial cells to certain extent.Compared with control group,the capacity of forming capillaries and VEGF secretion of serum deprivation groups enhanced significantly(P < 0.05).which validate the possibility of converting fibroblasts into endothelial cell with nongene-integration strategy for further optimization of IHD drug therapy of neovascularization of ischemic,at the same time,offerring theoretical basis for the mechanism research,clinical application,disease model,drug discovery for regenerative medicine foundation.Conclutions:Serum deprivation stimulation can activate cardiac fibroblasts to endothelial cell transdifferentiation,which suggests that fibroblasts-endothelial transition with without using gene marterial is feasible. |
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