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Expression of major blood group antigens on human erythroid cells in a two
phase liquid culture system
H Wada, T Suda, Y Miura, E Kajii, S Ikemoto and Y Yawata
Department of Medicine, Kawasaki Medical School, Kurashiki, Japan.
In order to examine the sequential expression of major blood group antigens
on human erythroblasts, a selective two phase liquid culture system for
erythroid progenitors was established. After mononuclear cells obtained
from peripheral blood were cultured in the presence of phytohemagglutinin
stimulated-leukocyte conditioned medium (PHA-LCM) for 7 days (the first
phase), nonphagocytic cells were recultured under hypoxic culture
conditions containing 30% fetal calf serum, 1% bovine serum albumin, 300
micrograms/mL transferrin and 2 U/mL recombinant erythropoietin (the second
phase). Mature (orthochromatic) erythroblasts were observed on day 4 of the
second phase, and reached 57.1 +/- 3.1% of total cells on day 8, followed
by the appearance of denucleated red cells, equivalent to mature red cells
in peripheral blood. Hemoglobin contents reached the level of 16.8 +/- 0.7
micrograms/10(6) cells on day 8. Flow cytometric analyses revealed that, on
day 3 of the second phase, cells became blood type M-positive,
corresponding to the maturation of erythroid cells. Regarding the
expression of ABH blood group antigens, a small number of blood type H-
positive cells were initially detected on day 0 of the second phase, while
blood type A-positive cells, which essentially were not observed on day 0,
increased gradually corresponding to the extent of erythroid maturation. In
the present system, Lewis and P1 blood group antigens were expressed at day
5 of the second phase, although autologous plasma was required to determine
the expression of Lewis blood group antigens. This culture system is
beneficial for studies on normal and abnormal human red cell membranes,
because the erythroid progenitors in human peripheral blood were used, and
a reasonable number of erythroid cells (0.5 to 1.5 x 10(7] was obtained
with good maturation.
Volume 75,
Issue 2,
pp. 505-511,
01/15/1990
Copyright © 1990 by The American Society of Hematology
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