Isolation and transplantation of allogeneic pulmonary endothelium derived from GFP transgenic mice
Isolation and transplantation of allogeneic pulmonary endothelium derived from GFP transgenic mice
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DOI：
10.1016/j.jim.2003.08.004
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发表时间：
2003-12-01
影响因子：
2.2
通讯作者：
Gerbitz, A
中科院分区：
文献类型：
--
作者：
Ewing, P;Wilke, A;Gerbitz, A
长期以来，从小鼠组织中分离原代内皮细胞一直是一个挑战，而且仍然是一项艰巨的任务。以转GFP基因的C57/BL6小鼠为供体，经DiI-Ac-低密度脂蛋白(LDL)染色后，采用流式细胞术分离肺内皮细胞。小鼠肺组织机械分离和酶消化后，贴壁细胞可用流式细胞仪快速染色和分选。分离的细胞表达CD31、MECA32和CD106等内皮细胞标志物，Isolectin B4染色阳性。在使用标准的内皮细胞生长介质进行50倍扩增后，细胞可以被移植到致死照射的同种异体宿主中，并在移植后24小时内在肺组织中使用荧光显微镜进行检测。(C)2003爱思唯尔B.V.保留所有权利。
The isolation of primary endothelial cells from murine tissues has long been a challenge and remains a difficult task. Using GFP transgenic C57/BL6 mice as donors, we describe a reliable method to isolate pulmonary endothelial cells by flow cytometry after staining with DiI-Ac-low density lipoprotein (LDL). After mechanical dissociation of murine lung tissue and enzymatic digestion, adherent cells can be quickly stained and sorted by flow cytometry. The isolated cells express endothelial cell markers such as CD31, MECA32 and CD106 and stained positive for Isolectin B4. After 50-fold expansion using standard endothelial growth media, cells could be transplanted into lethally irradiated allogeneic hosts and were detectable using fluorescence microscopy up to 24-h post-transplantation in pulmonary tissue. (C) 2003 Elsevier B.V. All rights reserved.