Culture Of Muscle Satellite Cells And Effect Of Myocardial Homogenate On Transdifferentiation Of Satellite Cells

[Objective]SCs was transplanted into the site of cardiac injury and induced in the new circumstance. The biological character of SCs may adjust a little, make conjunction with the cardiocyte and express some cardiac specific protein. Because it is complicated and influenced by many factors, such as electrical and mechanical stimulating, we design the experiment that SCs was cultured in the circumstance of myocardial homogenate and observe whether the biological character has ever changed.[Methods] SCs were digested with collagenase I and Trypsin,which were isolated and purified with discontinuous density centigugation(Percoll gradient liquid).After establishing stable culture methods for rats SCs and identification of SCs withα-Actin monoclonal antibody and SABC immunochemistry stains, they were induced by myocardial homogenate on day 3 of primary culture for 1 week .The morphological change and differentiated character of SCs were observed ,and Imumunocytochemistry technique was used to detect cardiac troponin-T(cTn-T) 3 weeks later.[Results] After SCs were digested with mixture of collagenase I and Trypsin, isolated and purified with discontinuous density centigugation, SCs purity could be as high as 90%.SCs look like spindles and fused into myotubes at the forth day of the culture phase. Byα-actin immunochemistry stains tests, Yellowish brown color particle was found in the cytoplasm. SCs of primary culture did not adapt in the new circumstance and die in a great quantity in the early phase. The immunochemistry stains test of cTnT proved negative 28 days later.[Conclusion]SCs of primary culture did not adapt in the new circumstance and die in a great quantity in the early period. The immunochemistry stains test of cTnT proved negative 28 days later. SCs is still a perspective seed cell, but need be treated before being transplated such as being pretreated with heat shock, or transfected with some helpful genes so that the survival rate of seed cells can increase , angiogenesis of the site of cardiac injury can increase and SCs can effectively conjunct with cardiocyte.