The Mechanism Of Exploration Of Stem Cells In Intrauterine Adhesion Anti-fibrosis

Objective Assess the anti-fibrosis action potential among umbilical cord mesenchymal stem cells(UCMSC),fallopian tube mucosa mesenchymal stem cells(FMMSC)and endometrial stem cells(En MSC),preliminary explore the mechanism of action of stem cell therapy caused by intrauterine adhesions anti-fibrosis,to seek a better stem cells for the treatment of Intrauterine adhesions anti-fibrosis.Methods To isolate,culture and identify UCMSC,En MSC and FMMSC in vitro.To observe the cell morphology by inverted microscope,using flow cytometry instrument to analyze the cell phenotype and cell cycle,flat cell clone formation test its proliferation,CCK 8 method to determine its growth characteristics,karyotype analysis to detect the genetic stability and differentiation in vitro experiments show multi-directional differentiation potential,Real Time polymerase chain reaction to detect UCMSC,En MSC and FMMSC anti-fibrosis related proteins relative to express.Results1.The three sources of different tissue cells inoculated in MSC complete medium,cells adherent about 12 h,cultivate to 5d,about 80% cell fusion.Cells were observed under inverted microscope in fifth generation,characterized by uniform long spindle,and swirl,UC source cells form more uniform,slender,closely packed,FM stereo sense is stronger,and En source cells form more similar to the form.2.Three types of MSC were expression of CD90 CD73,CD105,CD29,CD44,CD166,and HLA-ABC,lack expression of CD14,CD19,CD45,CD34 and HLA-DR.3.The cell cycle detection shown that the proportion of FMMSC was higher than UCMSC and En MSC in DNA synthesis period(G2 + S).4.Flat cell clone formation test,according to the results of clone groups of UCMSC was 136±26.32,FMMSC was 89±5.34,En MSC was 91.6±91.6,clone formation rates were 136%±26.32%,89%,±5.34% and 91.6%,±2.7%.UCMSC and FMMSC,En MSC cloning number and clone formation rates difference was statistically significant(P <0.01),but FMMSC and En MSC were no statistically significant difference.5.The CCK-8 growth curve was S-shaped.UCMSC latent time is shorter,about 24 h in logarithmic phase.FMMSC was entering exponential phase grow fast after 4 to 6 d,in the 7-8d into the plateau.6.Karyotype results conformed to the source of sample,the cells cultured by this method with the genetic stability.7.The induced differentiation experiments indicated that UCMSC,FMMSC and En MSC could be induced into osteoblast,adipocyte and chondrocytes.8.According to the results of Anti-fibrosis associated protein m RNA expression,FMMSC and En MSC HGF,IL-10 and IFN-gamma secretion more than UCMSC(P <0.05),while FMMSC and En MSC were indifference.Three groups of stem cells b FGF,Adrenomedullin and lefty secretion were no statistical difference,but there are differences in visual.The relative expression of b FGF,FMMSC and En MSC were higher than UCMSC.En MSC Adrenomedullin secretion was significantly more than FMMSC and En MSC.FMMSC lefty secretion than En MSC.En MSC lefty secretion than UCMSC.Conclusions1.The above FMMSC UCMSC proliferation ability and En MSC,but FMMSC proliferation faster;2.FMMSC and En MSC biological characteristics are highly similar;3.UCMSC,FMMSC and En MSC are secreted anti-fibrosis related protein;4.The proportion of anti-fibrosis effection in FMMSC and En MSC,HGF,IL-10 and IFN-gamma may be more than b FGF,Adrenomedullin and lefty.HGF,IL-10 and IFN-gamma involved in pathways may be play an important role of anti-fibrosis.5.In patients,autologous transplantation can be realized through using FMMSC.Compare UCMSC with FMMSC,as a better alternative stem cell,FMMSC has higher clinical value for the treatment of endometrial damage.