Modderman W E, Vrijheid-Lammers T, Löwik C W, Nijweide P J
Laboratory of Cell Biology and Histology, University of Leiden, The Netherlands.
Exp Hematol. 1994 Feb;22(2):194-201.
A method is described that permits the removal of hematopoietic cells and macrophages from mouse bone marrow cultures. The method is based on the difference in effect of extracellular ATP4- ions (ATP in the absence of divalent, complexing cations) on cells of hematopoietic origin, including macrophages, and of nonhematopoietic origin, such as fibroblastlike stromal cells. In contrast to fibroblastlike cells, hematopoietic cells and macrophages form under the influence of ATP4- lesions in their plasma membranes, which allows the entrance of molecules such as ethidium bromide (EB) and potassium thiocyanate (KSCN), which normally do not easily cross the membrane. The lesions can be rapidly closed by the addition of Mg2+ to the incubation medium, leaving the EB or KSCN trapped in the cell. This method allows the selective introduction of cell-toxic substances such as KSCN into hematopoietic cells and macrophages. By using this method, fibroblastlike stromal cells can be isolated from mouse bone marrow cultures.
本文描述了一种从小鼠骨髓培养物中去除造血细胞和巨噬细胞的方法。该方法基于细胞外ATP4-离子（不含二价络合阳离子的ATP）对造血起源细胞（包括巨噬细胞）和非造血起源细胞（如成纤维样基质细胞）作用的差异。与成纤维样细胞不同，造血细胞和巨噬细胞在ATP4-的影响下，其质膜会形成损伤，从而使溴化乙锭（EB）和硫氰酸钾（KSCN）等通常不易穿过细胞膜的分子得以进入。通过向孵育培养基中添加Mg2+，损伤可迅速闭合，使EB或KSCN被困在细胞内。该方法允许将细胞毒性物质如KSCN选择性地引入造血细胞和巨噬细胞。利用此方法，可从小鼠骨髓培养物中分离出成纤维样基质细胞。
