Comparative Expression Profiles Of Micrornas Between Adipose Tissue-Derived Mesenchymal Stem Cells And Myocardial Cells Induced From ADMSC

Objective:1.To explore the method for isolation and cultivation of Adipose Tissue-Derived Mesenchymal Stem Cells(ADMSCs).2.To prove that ADMSCs can be induced into myocardial cells by5-aza-2-deoxycytidine (5-Aza).3.To explore expression profiles of miRNAs between ADMSCs and Myocardial Cells Induced from ADMSC.Methods:1. Isolating, culturing and identifying ADMSCs. Under aseptic conditions, get inguinal fat tissue from SD rats, digest the tissue using trypsin and type I collagenase sift off and culture the collected cells; Collecting the third-generation cells for surface molecular identification.2.ADMSCs differentiation and identification in vitro:induced by5-Aza, the expression of a-actin, cTnT were examined by immunecytochemical method.3. Detect and quantify expression profiles of miRNAs:Three groups were detected.The third-generation ADMSCs were assigned into the first group.The cells cultured without5-Aza were assigned into the second group. The cells induced by5-Aza and cultured4weeks were assigned into the third group. MiRNA A-Sensor Array was used to detect and quantify expression profiles of miRNAs, and the profiles of miRNAs were compared.Result:1. There was a large amount of mesenchymal stem cells in SD rats adipose tissue. Immunocytochemical staining showed that most of cultured cells were CD13、CD44、 CD105positive, CD34、CD45、HLA-DR and factor Ⅷ negative. 2.Induced cells became bigger and longer. The connection of the cells was formed.The direction of cell arraying was gradually similar.After the5-Aza treatment, the cells were stained positively for a-actin and cTnT.3. There were35miRNAs which RIF (Relative Inhibiting Fold) were not less than5in first group. All the expression levels increased after induced by5-Aza.The most obviously15miRNAs were listed below:let-7a, let-7b, let-7e, let-7f, let-7g, miR-125a-5p, miR-142-5p, miR-143, miR-145, miR-21, miR-222, miR-34c-5p, miR-92a.Conclusion:1.SD rats fat tissue contains mesenchymal stem cells.2.ADMSCs induced by5-Aza may differentiate into myocardial cells.3.The method called MiRNA A-Sensor Array could detect and quantify expression profiles of a large number of miRNAs at the same time continually. This method could be used to monitor the miRNA activity in vitro.4.Many miRNAs play an important role in the process of cell differentiation,and the most important miRNAs may be let-7family and miR-21.