应用Percoll分离纯化组织工程用肌卫星细胞

Separation and Purification of Skeletal Muscle Satellite Cells for Tissue Engineering Applications by Percoll

目的 探讨应用改良Percoll非连续密度梯度离心法分离纯化培养组织工程用肌卫星细胞。方法 分别采用差速贴壁法和Percoll非连续密度梯度离心法进行兔肌卫星细胞的原代培养、光镜及扫描电镜检查细胞形态，通过免疫细胞化学染色方法鉴定培养细胞纯度，噻唑蓝法测定细胞生长曲线。结果 差速贴壁法所得细胞纯度为30％．40％，Percoll非连续密度梯度离心法所得细胞纯度在90％以上，两种方法光镜及电镜检查均符合卫星细胞形态，细胞生长曲线显示细胞生长状况良好。结论 应用Percoll非连续密度梯度离心法可以得到高纯度的肌卫星细胞，可以作为组织工程用种子细胞的培养方法。

Objective To investigate separation and purification of skeletal muscle satellite cells with improved incontinuous density Percoll gradient centrifugation technique. Methods The primary skeletal muscle satellite cells of New Zealand white rabbits were cultured with different adhesion time method and incontinuous density Percoll gradient centrifugation technique. The cells were observed under invert microscope and scanning electron microscope. The degree of purification was examined by celluar immunocbemical stain. The growth curve was tested by thiazolyl blue assay. Results Over 90% satellite cells were harvested by incontinuous density Percoll gradient centrifugation technique, in contrast to which, only 30% -40% cells were harvested by different adhesion time. Morphological observation accorded with satellite cells. The growth curve indicated that the cells growed in a good status. Conclusion The high purification satellite cells can be obtained by incontinuous density Percoll gradient centrifugation technique. It is a good method to culture seed cells for tissue engineering applications.