一种分离Wistar大鼠中幼、晚幼有核红细胞的有效方法

A HIGH EFFICIENT METHOD FOR THE SEPARATION OF CELL FRACTION ENRICHED WITH INTERMEDIATE AND LATE ERYTHROBLAST CELLS

本文介绍采用改良的Harrison分离骨髓造血细胞方法,用40%和70%不连续Percoll介质梯度,从15天Wistar鼠胚肝中分离中幼、晚幼有核红细胞。从70%Percoll梯度界面(比重1.095 g/ml)可收集到高达96%左右的中幼、晚幼有核红细胞。经Giemsa染色和联苯胺染色,光镜观察以及透射电镜检查鉴定,其中除极少量早幼红细胞和网织红细胞外,几乎没有其它类型细胞混杂。台盼蓝染色检查表明,分离后的中幼、晚幼红细胞保持95%以上的活性率,可用于细胞生物学和生物化学等分析。与其它同类方法比较,本方法有取材容易、操作迅速、简便和获取量高等优点。

A method described by Harrison has been adopted and modified by us for the separation of intermediate and late erythroblast cells from 15 day embryonic liver of pregnant Wistar rat.The method consisted briefly of preparation of fetal liver cell suspension and the separation of cell types in a 40%and 70%nonlinear Percoll gradient system.Using this method,we can obtain about 96%of hemogenous population of intact and viable intermediate and late erythroblasts.Examination of tho separated cells by Giemsa and benzidine staining and by electron microscopic observation indicated that no granulocytes or other white blood cells could be detected,except for some contamination of about 1%of proerythroblasts and 3%of reticulocytes in the fraction.Trypan blue vital staining demonstrated that there were over 95%of the cells maintained viable after separation,they could be used directly for the study of cell differentiation as well as biochemical analysis.SO,this is an economical,simple and easy technique to operate which proved to be a useful mean for obtaining enrich population of intermediate and late erythroblasts in research in the field of cell biology.