Study Of Respectively Co-culture Of Human Amniotic Mesenchymal Stem Cells And Human Umblical Cord Stem Cells With C6 Glioma Cells

Background and Objective:Glioma is the most common intracranial tumor,although the elements in glioma, the study of a lot of progress has been made,but the prognosis of the patients are still not satisfied.At present surgical resection is still the preferred method of treatment, but the radical resection of the high relapse rate after the impact of its long-term survival rate is the main factor.Recently,we can fine some reports about the MSCs can depress the growing of the tumor.Our team have proved that MSCs can depress the growing of the tumor stem cells during the previous experiments.Because the C6 golima cells can be get easily and stably,so the basic research about C6 golima cells can enrich the knowledge of golima.This experiment in the establishment of hMSCs and hUSCs in vitro culture and identification methods on the basis of hMSCs and hUSCs to observe the probably different result which can be get by different method of co-culture between the MSCs and C6 golima cells.In this study,we will carry out in vitro seperatly with the method of co-culture between the MSCs and C6,initially found that the deferent interaction between the MSCs and C6,further defined in the choice ang reference of the next research. Materials and Methods:Under sterile conditions for the normal full-term fetus amniotic caesarean section and umlilicalcord,refine hMSCs and hUSCs from them, culture in containing 10%fetal bovine serum(fetal bovine serum,FBS) DMEM/F12 the culture medium with the method of explant tissue and digestion.P3 hMSCs and hUSCs from the immunohistochemistry and flow cytometry analysis:CD29,CD44 positive expression;flow cytometry test cycle with stem cell characteristics;P3 generation hMSCs and hUSCs co-culture with C6 directly and indirectly,there will be 5 groups,Direct train between hMSCs and C6(Group A),Indirect train between hMSCs and C6(Group B),Direct train between hUSCs and C6(Group C),Indirect train between hUSCs and C6(Group D),Blank control of C6(Group E),collected a total of cultured C6 for flow cytometry determination and electron-microscopic analysis.Results:Digestive enzymes separation of amniotic MSCs,can be cultivated in vitro and then through the wall of separation has been continuously purification.Digestive enzymes and explant tissues separation of umbilicalcord MSCs,can be cultivated in vitro and then through the wall of separation has been continuously purification.Immunocytochemical staining showed that CD44 and CD29of hMSCs and hUSCs are positive;the cell cycle analysis of hMSCs and hUSCs with stem cell characteristics of a typical cycle,FCM passage hMSCs negative control 9.39%,CD29 positive cells ratio was 82.53%,CD44 positive cells ratio was 90.86%,HLA-ABC positive cells ratio was 89.55 percent,hUSCs negative control7.61%,CD29 positive cells ratio was 70.44%,CD44 positive cells ratio was 75.50%.Under the electron-microscopic,we can find that all the four groups show the jionts tail off,the volume tail off,cellural organ denaturation,even the apoptotic body can be find,Group E grow up vigorously.Group A,B,C,D show that the bcl-2 postive rate of C6 reduce with the time run out,and the Group C,D will be more remarkable tha Group A,B;Group A,Cwill be more remarkable tha Group A,B;Group A,B,C,D show that the bcl-2 postive rate of C6 reduce with the time run out,and the Group C,D will be more remarkable tha Group A,B;Group A,Cwill be more remarkable tha Group A,B.Conclusions:1.Bcl-2 in C6 was in high expression.2.hMSCs and hUSCs can reduce C6-cell cloning between the role of adhesion.3.C6 proliferation weakened in co-culture with hMSCs and hUSCs.4.Bcl-2 in C6 after the co-culture will be lower with timing flow.5.The apoptosis rate of C6 after the co-culture will be upper with timing flow.6.Indirect co-culture group of C6 depressed more extremly.7.hUSCs depress the growing of C6 more extremly.