Cajal间质细胞的分离、培养方法探讨

Isolation and culture of interstitial cells of Cajal

目的 探讨体外培养Cajal间质细胞 (interstitialcellsofCajal,ICC)的方法。 方法 以小鼠小肠为ICC的组织来源 ,应用酶解法并结合密度离心法分离、培养ICC ,对ICC培养中的污染、Ca2 + 、Mg2 + 离子以及胶原酶等影响因素进行了探讨 ,应用ICC特异的c kit抗体进行免疫荧光染色后在激光共聚焦显微镜对其进行了鉴定。结果 成功建立起一套的分离、培养的方法。免疫荧光鉴定结果提示 ,本方法建立的ICC分离培养方法是稳定可靠的。结论 ICC分离培养比较困难 ,需要注意防止污染、减少无Ca2 + 、Mg2 + 离子液体以及胶原酶消化时间等问题 ,本方法有望为致力于ICC细胞研究的科研工作者提供方法借鉴 ,并有望促进ICC研究的深入进行。

Objective To discuss the method for isolation and culture of interstitial cells of Cajal(ICC). Methods ICC were isolated from the ileal segment of mice. Enzymatic dissociation and Ficoll density centrifugation were used to culture ICC. Factors effecting the successful culture of ICC, such as contamination, the solution without Ca 2+ , Mg 2+ and collagenase were investigated during the study. The cultured ICC were stained with c kit immunofluorescence antibody and identified under the confocal microscopy. Results We have successfully built up the method for the culture of ICC. Conclusions Many factors which are essential for the culture of ICC should be mentioned, such as preventing contamination, reducing exposure to solution without Ca 2+ , Mg 2+ and appropriate collagenase digestion.