In Vitro Culture of Erythroid Colonies from Human Fetal Liver and Umbilical Cord Blood

Abstract

Summary. 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 10⁶ 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 seen 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 β and γ 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.