Mitjavila M T, Natazawa M, Brignaschi P, Debili N, Breton-Gorius J, Vainchenker W
INSERM U 91, Hôpital Henri Mondor, Créteil, France.
J Cell Physiol. 1989 Mar;138(3):617-23. doi: 10.1002/jcp.1041380324.
Erythroid progenitors from normal human marrow were purified by a two-step immune panning method permitting both the enrichment of erythroid progenitors (plating efficiency up to 10%) and the separation of CFU-E from BFU-E. The purified erythroid progenitors were grown in serum-replaced conditions; in some experiments at an average of one cell per well. Human recombinant granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin 3 (IL3), erythroid potentiating activity (EPA), and human erythropoietin (Epo) either recombinant or homogenous native were tested for their effect on CFU-E growth. Epo was an absolute requirement for CFU-E growth and was sufficient to obtain colony formation at the unicellular level whereas GM-CSF and IL3 did not further increase the plating efficiency. EPA potentiated the effect of Epo on this progenitor only in experiments performed at unicellular level. Human recombinant GM-CSF, IL3, Interleukin 1 alpha (IL1 alpha), and Epo were subsequently tested for their ability to promote BFU-E growth. GM-CSF and IL3 supported the growth of erythroid bursts in the presence of Epo, even at the unicellular level. However, IL3 promoted a higher number of bursts than GM-CSF under all conditions tested. These two growth factors have no or very small additive effects when tested in combination. IL1 alpha added to Epo alone had no effect on the growth of BFU-E whereas it potentiated the combined action of IL3 and GM-CSF on the primitive BFU-E. In conclusion, this study confirms at the unicellular level and under serum-free conditions that erythroid progenitors are regulated by multipotential growth factors in early phases of erythropoiesis and become sensitive only to Epo in later phases of differentiation.
采用两步免疫淘选法对正常人骨髓中的红系祖细胞进行纯化，该方法既能富集红系祖细胞（铺板效率高达10%），又能将CFU-E与BFU-E分离。纯化后的红系祖细胞在无血清条件下培养；在一些实验中，平均每孔接种一个细胞。对重组人粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素3（IL3）、红系增强活性（EPA）以及重组或同源天然的人促红细胞生成素（Epo）对CFU-E生长的影响进行了测试。Epo是CFU-E生长的绝对必需因子，足以在单细胞水平获得集落形成，而GM-CSF和IL3并未进一步提高铺板效率。仅在单细胞水平进行的实验中，EPA增强了Epo对该祖细胞的作用。随后对重组人GM-CSF、IL3、白细胞介素1α（IL1α）和Epo促进BFU-E生长的能力进行了测试。即使在单细胞水平，GM-CSF和IL3在有Epo存在的情况下也支持红系集落的生长。然而，在所有测试条件下，IL3促进形成的集落数量均多于GM-CSF。这两种生长因子联合测试时无相加作用或相加作用非常小。单独添加到Epo中的IL1α对BFU-E的生长无影响，而它增强了IL3和GM-CSF对原始BFU-E的联合作用。总之，本研究在单细胞水平和无血清条件下证实，红系祖细胞在红细胞生成早期受多能生长因子调控，而在分化后期仅对Epo敏感。
