Hassan M W, Lutton J D, Levere R D, Rieder R F, Cederqvist L L
Br J Haematol. 1979 Apr;41(4):477-84. doi: 10.1111/j.1365-2141.1979.tb05885.x.
Colony formation by erythroid precursors from human fetal liver, umbilical cord blood and adult peripheral blood has been studied in a plasma clot culture system. Fetal liver (FL) was obtained at post-mortem examination from 13-22 week abortuses. After mincing in Hanks' solution, cells in suspension were harvested by Ficoll-Hypaque centrifugation. Mononuclear cells were obtained by centrifugation of umbilical cord blood (CB) and normal adult peripheral blood (PB). All three types of preparations were incubated up to 14 d in 0.1 ml plasma clot cultures containing 0-4 u/ml erythropoietin (Epo) and 10(6) cells/ml. No colonies formed in the absence of Epo. Normal adult PB produced late-appearing colonies; there were no colonies at day 7 and up to 100 colonies/0.1 ml at day 14. CB produced early and late colonies with up to 200 colonies/0.1 ml at day 7 and 125 at day 14. Cells from FL produced many early colonies; over 1500 colonies/0.1 ml were sent at day 7 and there was a subsequent decline in colony count with longer incubation. In cultures of both CB and FL, colonies composed of either mature or immature cells were noted during both early and late stages of incubation suggesting that these cell sources contain a heterogeneous population of erythroid colony progenitors. Measurement of differential beta and gamma globin chain synthesis by erythroid colonies grown from fetal liver and umbilical cord blood gave results similar to those obtained by direct pulse-labelling of the original source of the cultured cells.
在血浆凝块培养系统中,对来自人类胎儿肝脏、脐带血和成人外周血的红系前体细胞的集落形成进行了研究。胎儿肝脏(FL)是在对13 - 22周流产胎儿进行尸检时获得的。在汉克斯溶液中切碎后,通过菲可-泛影葡胺离心收集悬浮细胞。通过对脐带血(CB)和正常成人外周血(PB)进行离心获得单核细胞。所有三种类型的细胞制剂在含有0 - 4 U/ml促红细胞生成素(Epo)和10⁶个细胞/ml的0.1 ml血浆凝块培养物中孵育长达14天。在没有Epo的情况下不形成集落。正常成人PB产生出现较晚的集落;在第7天没有集落,在第14天可达100个集落/0.1 ml。CB产生早期和晚期集落,在第7天可达200个集落/0.1 ml,在第14天为125个。来自FL的细胞产生许多早期集落;在第7天发出超过1500个集落/0.1 ml,随着孵育时间延长集落数量随后下降。在CB和FL的培养物中,在孵育的早期和晚期都观察到由成熟或未成熟细胞组成的集落,这表明这些细胞来源包含异质性的红系集落祖细胞群体。对从胎儿肝脏和脐带血生长的红系集落的β和γ珠蛋白链合成差异的测量结果,与通过对培养细胞的原始来源进行直接脉冲标记获得的结果相似。