The Preliminary Study Of Therapeutic Mechanism Of Glucocorticoid By Injection Into Sternocleidomastoid Muscle In CMT

[objective]In order to establish the method for primary culture of myoblast derived from human normal skeletal muscle and contractural muscle and to prepare for the following experiment.[Methods]The first part: The skeletal muscularsamples of 4-7 days rats were minced and digested by collagenaseI and purified with Percoll gradient liquid and trypsin . .The cells were then purified by differential attachment.We observe not only their proliferation and differentiation characteristic under the contrast phase microscope but the forming process of the polynuclear myotube.The second part: The skeletal muscular samples derived from human normal muscle after operation were isolated and cultured by the method of trypsin and collagenase I digestion. These kinds of cells were tested by actin immunohistochemical stains. We also observe their proliferation and differentiation characteristic under the contrast phase microscope and the forming process of the polynuclear myotube.The third part: The skeletal muscular samples derived from contractural muscle of congenital muscular tortiocllis patients after operation were isolated and cultured by the method of trypsin and collagenase I digestion.[result]1. We can observate the forming process of the polynuclear myotubes at 4th and 5th day after the cell planted in first part of experiment.They have many cell nucleuses which arrange by the string of beads type.2. We can also observate the forming process of the polynuclear myotubes at 13th and 14th day after the cell planted in the second part of experiment.They have many cell nucleuses which arrange by the string of beads type. Cultured satellite cells derived from human normal muscle after operation diferentiated into myoblasts and fused to form myotubes later.These cells were positive by actin antibody's staining.3. We have not successfully cultured myoblast from contractural muscle of congenital muscular tortiocllis patients after operation.[conclusion]A large number of myoblasts can be obtained from normal muscle of CMT patient after operation by enzyme digestion and repeated attachment method. The cultured cells can be proved as myoblasts by morphological and immunohistochemical detection.However,we have not successfully cultured myoblast from contractural muscle of congenital muscular tortiocllis patients after operation according to present method.