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肥大细胞生长因子调节人骨髓和外周血中与持续分化相关的红系祖细胞上CD36抗原的表达。

Mast cell growth factor modulates CD36 antigen expression on erythroid progenitors from human bone marrow and peripheral blood associated with ongoing differentiation.

作者信息

de Wolf J T, Muller E W, Hendriks D H, Halie R M, Vellenga E

机构信息

Department of Hematology, University Hospital Groningen, The Netherlands.

出版信息

Blood. 1994 Jul 1;84(1):59-64.

PMID:7517219
Abstract

To study the differentiation process of erythroid progenitors from normal human bone marrow and peripheral blood, CD34/CD36 sorted cells were cultured in the presence of Erythropoietin (Epo) and Epo plus mast cell growth factor (MGF). The CD34+/CD36- cell fraction from bone marrow supported 74 +/- 33 erythroid burst forming units (BFU-E)/10(4) cells (mean +/- SD, n = 4) in the presence of Epo, which increased 2.1-fold by coculturing with MGF. However, erythroid colony-forming units (CFU-E) were not cultured from the CD34+/CD36- cell fraction. In contrast, the CD34-/CD36+ cell fraction supported CFU-Es in the presence of Epo (152 +/- 115/10(5)) or Epo plus MGF (180 +/- 112/10(5)), whereas BFU-Es were hardly noticed. However, the transition of the BFu-E to CFU-E was observed by incubating CD34+/CD36- cells (10(4)/100 microL) in suspension with Epo plus MGF for 7 days followed by Epo in the colony assay. This was reflected by the appearance of CD34-/CD36+/Glycophorin A+/CD14- cells. In addition high numbers of CFU-Es (1,000 +/- 150, n = 4) were cultured from this cell fraction. In contrast to bone marrow erythroid progenitors, no peripheral blood CFU-Es were cultured from either the CD36+ or CD36- fraction, whereas BFU-Es were predominantly present in the CD36+ fraction. However, the CD34+ progenitor cell from peripheral blood did have intrinsic capacity to differentiate to CFU-Es because CD34+/CD36- cells incubated with Epo plus MGF for 7 days and followed by Epo in the colony assay, supported high numbers of CFU-Es (1,200 +/- 400, n = 3). To study whether additional growth factors have similar effects on erythroid progenitors, experiments were performed with interleukin 1 (IL-1), IL-3, and IL-6. IL-1 and IL-6 did not modulate the Epo supported proliferation and differentiation. In contrast, IL-3 in the presence of Epo did support CFU-Es, from CD34+/CD36- cells after 7 days in suspension culture. However, flow cytometry analysis showed that Epo plus IL-3 not only supported CD34-/CD36+/Glycophorin A+ cells but also CD36+/CD14+ cells, indicating the differentiation along different cell lineages. In summary, the data show a phenotypic distinction between bone marrow and peripheral blood erythroid progenitors with regard to CD36 expression. In addition, the results suggest that Epo plus MGF or IL-3 and preincubation in suspension culture are prerequisites for the transition of the BFU-E to the CFU-E.

摘要

为研究正常人骨髓和外周血中红系祖细胞的分化过程,将经CD34/CD36分选的细胞在促红细胞生成素(Epo)以及Epo加肥大细胞生长因子(MGF)存在的条件下进行培养。在Epo存在时,骨髓来源的CD34⁺/CD36⁻细胞亚群每10⁴个细胞可支持74±33个红系爆式集落形成单位(BFU-E)(均值±标准差,n = 4),与MGF共培养后增加2.1倍。然而,CD34⁺/CD36⁻细胞亚群未能培养出红系集落形成单位(CFU-E)。相反,CD34⁻/CD36⁺细胞亚群在Epo存在时(152±115/10⁵)或Epo加MGF时(180±112/10⁵)可支持CFU-E形成,而BFU-E几乎未被检测到。然而,通过将CD34⁺/CD36⁻细胞(10⁴/100 μL)与Epo加MGF悬浮培养7天,随后在集落测定中加入Epo,可观察到BFU-E向CFU-E的转变。这表现为CD34⁻/CD36⁺/血型糖蛋白A⁺/CD14⁻细胞的出现。此外,从该细胞亚群培养出了大量CFU-E(1000±150,n = 4)。与骨髓红系祖细胞不同,外周血中无论是CD36⁺还是CD36⁻亚群均未培养出CFU-E,但BFU-E主要存在于CD36⁺亚群中。然而,外周血来源的CD34⁺祖细胞具有分化为CFU-E的内在能力,因为将CD34⁺/CD36⁻细胞与Epo加MGF培养7天,随后在集落测定中加入Epo,可支持大量CFU-E形成(1200±400,n = 3)。为研究其他生长因子对红系祖细胞是否有类似作用,使用白细胞介素1(IL-1)、IL-3和IL-6进行了实验。IL-1和IL-6未调节Epo支持的增殖和分化。相反,在悬浮培养7天后,Epo存在时IL-3确实可支持CD34⁺/CD36⁻细胞形成CFU-E。然而,流式细胞术分析显示,Epo加IL-3不仅支持CD34⁻/CD36⁺/血型糖蛋白A⁺细胞,还支持CD36⁺/CD14⁺细胞,表明细胞沿着不同细胞谱系分化。总之,数据显示骨髓和外周血红系祖细胞在CD36表达方面存在表型差异。此外,结果表明Epo加MGF或IL-3以及悬浮培养预孵育是BFU-E向CFU-E转变的前提条件。

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