The study of biological characteristics of myoblasts co-cultured with Schwann cells / 中华骨科杂志

ABSTRACT

Objective In order to understand the relationship between myoblast and Schwann cell,our purpose was to investigate the effects of biological characters of myoblasts co-cul tured with Schwann cells and pro vide the basic theory for constructing artificial muscle involving artificial nerve.Methods After sterilizing with iodine tincture and alcohol,the brachial plexus,sciatic nerve and triceps surae muscle of neonatal SD rat were harvested and peeled off their membranes,blood vessels and fat tissues under operating microscope thor oughly.The nerve and muscle tissues were cut in pieces by microscis sors,and then digested and isolated by collagenase and pancreatin in20and15minutes respectively.DMEM medium was employed to culture my oblasts and Schwann cells.After co-culturing myoblasts of rats with Schwann cells in vitro,the morphological characteristics and the growth condition of two cells were observed under inverted phase contrast mi croscope,the effects of Schwann cells secretion for proliferation of myoblasts were detected by 3 HTdR iso topic tracing and expressed by disintegration per minute(DPM),formation rate of myotubes was counted under micro scope and statistic data was analyzed,the functional differentiation degree of my oblasts affected by Schwann cells was analysed by?-sarcomeric actin immunohistochemistry(SABC)and imaging analysis tech nique.Results Co-cultured myoblasts proliferated,and myotubes ap peared earlier.Comparing with sole-cultured my oblasts,the shape of myobutes from co-cultured myoblasts tended to be elongating and robust.The value of DPM far-exceeded the control group,and reached its peak of 2500(just800for con-trol group).The positive cells of ?-sarcomeric actin appeared in brown red color.However,syn thesis and excre tion of a-sarcome ric actin in co-cultured myoblasts were much greater than control group,and the gray ash value between two groups was of a significant difference.Conclusion Primary rat myoblasts co-cultured with Schwann cells in vitro is beneficial in regulating its the growth,proliferation and the differentiation.