大鼠CD4^+CD25^+调节性T细胞的分选和提取

Isolation and extraction of rat CD4^+CD25^+ regulatory T cells in vitro

目的分选提取大鼠脾脏和淋巴结CD4+CD25+调节性T细胞并进行纯度和活性检测。方法用免疫磁珠两步法既阴性选择和阳性选择分选提取大鼠CD4+CD25+调节性T细胞,流式细胞仪检测细胞纯度,台盼蓝染色计算细胞存活率。结果免疫磁珠分选提取的CD4+CD25+调节性T细胞纯度在86%以上,并且活性良好,活细胞百分率大于97%。结论通过免疫磁珠分选能够提取高纯度高活性的CD4+CD25+调节性T细胞。

Objective： To isolate and extract the rat spleen and lymph nodes CD4^＋ CD25^＋ regulatory T cells and detect the purity and activity.Methods： The isolation of rat CD4^＋CD25^＋ regulatory T cells was done using a two-step procedure.CD4^＋ T cells were initially isolated by negative selection,and CD25＋ T cells were then isolated by positive selection from the CD4＋ T cell fraction.The purity was analyzed by flow cytometry（FCM）.Trypan blue was stained to calculate the cell survival rate.Results： The purity of CD4^＋CD25^＋ regulatory T cells was more than 86% with the survival rate of over 97%.Conclusion： Immunomagnetic cell sorting（MACS） can extract high purity and activity of CD4^＋ CD25^＋ regulatory T cells.