小鼠肝脏特络细胞的分离与鉴定

Isolation and identification of mouse hepatic telocytes

[目的]基于已有的特络细胞分离方法,构建一种更为稳定高效的肝脏特络细胞分离方法,为后续特络细胞对肝脏潜在调控作用的研究建立坚固的理论基础和实验支持.[方法]选取C57BL/6雄性野生小鼠;采用酶液多次短期消化及离心法分离肝脏特络细胞,并进行常规细胞培养;采用CD34+Vimentin免疫荧光染色鉴定所得细胞表型.[结果]成功分离了小鼠肝脏特络细胞;小鼠肝脏特络细胞在体外培养时呈CD34及Vimentin双阳性,肝星状细胞系呈CD34阴性及Vimentin阳性.[结论]本分离法可以获得优质的肝脏特络细胞,满足后续研究的需要.

[Objective] To develop an optimized method which is more stable and efficient in hepatic telocytes separation based on current method, and to establish a solid theoretical foundation and experimental support for further study of potential regulation of hepatic cells by telocytes. [Methods] C57BL/6 male wild mice were selected for isolation of liver telocytes through multiple short enzymatic digestions and centrifugation. CD34＋Vimentin immunofluorescence staining was used to identify the cell phenotype. [Results] Hepatic telocytes were successfully separated. Hepatic telocytes were both CD34 and Vimentin positive in vitro, while hepatic stellate cell line was CD34 negative and Vimentin positive. [Conclusion] High-quality telocytes were obtained through this optimized method and could be used for other studies.