Methodology of separation, purification, cultivation and  identification of rat bone marrow macrophage

LI Jing; et al

doi:10.13406/j.cnki.cyxb.2003.04.009

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Methodology of separation, purification, cultivation and identification of rat bone marrow macrophage

更新时间：2024-01-17

- Methodology of separation, purification, cultivation and identification of rat bone marrow macrophage
- Journal of Chongqing Medical University Issue 4, Pages: 436-439(2003)
- 作者机构：
  
  1. 重庆医科大学基础医学院组胚教研室
  2. 重庆医科大学基础医学院组胚教研室,重庆,400016
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2003.04.009
  CLC： R329.2
- Published：2003，
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[1]李静,王亚平.大鼠骨髓巨噬细胞的分离,纯化,培养以及鉴定[J].重庆医科大学学报,2003(04):436-439.

LI Jing, et al. Methodology of separation, purification, cultivation and identification of rat bone marrow macrophage. [J]. Journal of Chongqing Medical University (4):436-439(2003)
[1]李静,王亚平.大鼠骨髓巨噬细胞的分离,纯化,培养以及鉴定[J].重庆医科大学学报,2003(04):436-439. DOI： 10.13406/j.cnki.cyxb.2003.04.009.

LI Jing, et al. Methodology of separation, purification, cultivation and identification of rat bone marrow macrophage. [J]. Journal of Chongqing Medical University (4):436-439(2003) DOI： 10.13406/j.cnki.cyxb.2003.04.009.

摘要

目的 :为深入研究骨髓巨噬细胞的生物学功能

探索一种简便的骨髓巨噬细胞的分离

纯化和培养的方法。方法 :分离获取大鼠骨髓细胞

在 6 0 %DMEM培养液

2 0 %马血清

2 0 % （v/v）L92 9培养上清的诱导条件下进行体外贴壁培养

6～ 7天时获得纯度较高的贴壁细胞。采用倒置显微镜下观察生活状态、Wright′s染色光镜观察、电子显微镜观察检测贴壁细胞形态学 ;酸性磷酸酶及非特异性酯酶染色测定细胞内酶的表达 ;吞噬鸡红细胞及墨汁颗粒实验检测其吞噬功能 ;免疫细胞化学染色观察细胞表面标志等生物学技术鉴定培养的贴壁细胞性质。结果 :获得高纯度的巨噬细胞

且具有良好的吞噬功能

并且具备巨噬细胞的形态特征

特有的水解酶类及特有的表面标志 -CD68。结论 :本法是一种简易实用的体外分离、纯化、培养骨髓巨噬细胞的方法。

Abstract

Objective:To study the biological functions of bone marrow macrophage (BMM Φ) and to establish the methodology of separation

purification

cultivation and identification of BMM Φ.Methods:Using the techniques of anchorage-dependent culture of separated rat bone marrow cell (rBMC) in DMEM culture media (contain 20% horse serum

20% (v/v)L 929 conditioned media) in vitro

a lot of purified anchor cells were obtained

and these cells were identified with specifically biological marker of macrophage

such as 1

morphologicalobservation:invert phase contrast microscopy

light and electron microscopy;2

enzyme cytochemistry:acid phosphatase (ACP)

α-acetic acid naphthol esterase (α-ANE);3

phagocytic experiment:phagocytosis of chicken erythrocytes and prepared Chinese ink;4

immunocytochemistry: surface specific antigen of macrophage ( CD 68 stain).Results:The cells were purified having functional satisfactory macrophage according to morphological observation

enzyme cytochemistry

phagocytic experiment and immunocytochemistry.Conclusion:This is a simple and easy method for separation

purification

cultivation and identification of rat marrow macrophage.

关键词

骨髓巨噬细胞分离纯化鉴定

Keywords

Bone marrow macrophageseparationpurificationidentification

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