SD大鼠乳鼠咀嚼肌成肌细胞的原代培养

Primary Culture of Myoblast of Neonatal SD Rat’s Masticatory Muscle in Vitro

目的 建立SD大鼠乳鼠咀嚼肌成肌细胞体外培养方法。方法 分离SD大鼠乳鼠咀嚼肌,采用胰蛋白酶 和胶原酶多次消化法分离细胞,再用差速贴壁法对细胞进行纯化,所得的细胞进行Desmin免疫组化鉴定。结果 95%的培养的细胞Desmin胞浆呈阳性反应,台盼蓝检查细胞成活率超过94%。结论 多次消化和差速贴壁法可 获得高产量高纯度的成肌细胞。

Objective To establish the method for primary culture of myoblast of neonatal Sprague-Dawley rat's masticatory muscle in vitro.Methods The masticatory muscle of neonate mice was separated and digested by collagenase and trypsin.The cells were then purified by differential attachment.The cells were primarily cultured in vitro with F-12 culture liquid and its protein desmin was detected by immunocytochemistry.Results Immunohistochemistry analysis showed that 95% cell were stained positive for desmin which was mostly distributed in cytoplasm,and the cell viability were up to 94% by Trypan blue stain.Conclusion High quality and quantity myoblast can be obtained by repeated digestions and differential attachments.