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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。
This article describes the use of a modified Harrison method for the isolation of bone marrow hematopoietic cells from 40-day and 70% discontinuous Percoll media gradients to separate young and late-stage young erythroblasts from 15-day-old Wistar rat embryonic liver. Up to 96% of juvenile and late-juvenile erythroblasts can be collected from a 70% Percoll gradient interface (specific gravity 1.095 g / ml). Giemsa staining and benzidine staining, light microscopy and transmission electron microscopy were performed. Among them, almost no other types of cells were mixed except for very few erythroblasts and reticulocytes. Trypan blue staining showed that the separated young and old red blood cells maintained more than 95% activity, which can be used for cell biology and biochemistry analysis. Compared with other similar methods, the method has the advantages of easy drawing, rapid operation, simplicity and high availability.