Nijhof W, Wierenga P K
J Cell Biol. 1983 Feb;96(2):386-92. doi: 10.1083/jcb.96.2.386.
Erythroid progenitor cells, CFU-E (colony-forming-unit-erythroid), were isolated to practical homogeneity by a combination of three enrichment procedures. CFU-E were generated in large amounts in spleens of mice previously bled and treated with the erythropoiesis-suppressing drug thiamphenicol. The average CFU-E concentration in spleens from mice 4 d after the thiamphenicol-treatment was 10%. These CFU-E were separated from lymphocytes, erythrocytes, and granulocytes and their progenitor cells by centrifugal elutriation and Percoll density gradient centrifugation. A three- to five-fold enrichment was obtained by elutriation, leading to a CFU-E concentration of 45%. With the Percoll gradient another twofold enrichment was achieved, providing us with a 80-100% CFU-E cell population. The overall recovery of CFU-E was 60-70%. This is a cheap, rapid, and highly efficient method of obtaining large quantities of viable CFU-E. The sequential formation of two-, four-, and eight-cell colonies from CFU-E cultured in vitro was studied. These cells enable us to study the biochemical changes occurring in the differentiation process of an erythroid progenitor cell induced by the hormone erythropoietin. The morphological and some physical and biological properties of these cells are presented.
通过三种富集程序相结合,将红系祖细胞(集落形成单位-红系,CFU-E)分离至实际的均一状态。在先前放血并用红细胞生成抑制药物甲砜霉素处理的小鼠脾脏中大量产生CFU-E。甲砜霉素处理后4天,小鼠脾脏中CFU-E的平均浓度为10%。通过离心淘析和Percoll密度梯度离心将这些CFU-E与淋巴细胞、红细胞、粒细胞及其祖细胞分离。通过淘析获得了三到五倍的富集,使CFU-E浓度达到45%。利用Percoll梯度又实现了两倍的富集,为我们提供了80%-100%的CFU-E细胞群体。CFU-E的总体回收率为60%-70%。这是一种获得大量有活力CFU-E的廉价、快速且高效的方法。研究了体外培养的CFU-E依次形成二细胞、四细胞和八细胞集落的情况。这些细胞使我们能够研究由促红细胞生成素诱导的红系祖细胞分化过程中发生的生化变化。本文介绍了这些细胞的形态以及一些物理和生物学特性。