ISOLATION AND FLOW CYTOMETRIC ANALYSIS OF THE FREE LYMPHOMYELOID CELLS PRESENT IN MURINE LIVER
ISOLATION AND FLOW CYTOMETRIC ANALYSIS OF THE FREE LYMPHOMYELOID CELLS PRESENT IN MURINE LIVER
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DOI：
10.1016/0022-1759(90)90407-m
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发表时间：
1990-08-28
影响因子：
2.2
通讯作者：
MILON, G
中科院分区：
文献类型：
--
作者：
GOOSSENS, PL;JOUIN, H;MILON, G
在感染期间，肝脏中循环淋巴骨髓样细胞的募集通常在介导病原体生长的控制或恶化中起关键作用。本文介绍了一种简单而快速的技术，以恢复这些淋巴骨髓样细胞从正常或感染的肝脏。门静脉灌注生理盐水缓冲液后，肝脏在钢筛上轻轻解离，所得细胞群在35% Percoll 100 IU/ml Calciparine中离心，以去除所有细胞核和细胞碎片：通常在40-60 min内恢复纯肝淋巴髓样细胞群。然后可以轻松地对该细胞群进行表型和功能分析。将该方法应用于正常小鼠肝脏：纯化的游离淋巴骨髓样细胞的流式细胞术分析显示存在T淋巴细胞（46% ± 10%）。3，CD 4/CD 8比值为2.8），B淋巴细胞（20% ±. 2 IgG和30%IgM阳性）和骨髓单核细胞（14% ± 10%）。2例Ⅱ型补体受体阳性）。
Recruitment of circulating lymphomyeloid cells in the liver during infection often plays a critical role mediating control or exacerbation of the pathogen growth. This paper describes a simple and rapid technique to recover these lymphomyeloid cells from a normal or an infected liver. After portal perfusion with saline buffer, the liver is gently dissociated on steel screens and the resulting cell population spun in 35% Percoll in 100 IU/ml Calciparine to remove all nuclei and cell debris: the recovery of a pure liver lymphomyeloid cell population is usually achieved in 40-60 min. Phenotypic and functional analysis could then be easily carried out on this cell population. This methodology was applied to normal mouse liver: flow cytometric analysis of the purified free lymphomyeloid cells showed the presence of T lymphocytes (46% .+-. 3 with a CD4/CD8 ratio of 2.8), B lymphocytes (20% .+-. 2 IgG and 30% IgM positive) and myelomonocytic cells (14% .+-. 2 complement receptor type II positive).