The effect of thrombopoietin on the proliferation and differentiation of murine hematopoietic stem cells
The effect of thrombopoietin on the proliferation and differentiation of murine hematopoietic stem cells
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DOI：
10.1182/blood.v87.12.4998.bloodjournal87124998
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发表时间：
1996-06-15
期刊：
影响因子：
20.3
通讯作者：
Kaushansky, K
中科院分区：
文献类型：
--
作者：
Sitnicka, E;Lin, N;Kaushansky, K
在这项研究中，我们探讨了血小板生成素（Tpo）是否对长期再生的造血干细胞（LTR-HSC）的增殖和分化有直接的体外影响。我们之前报道了一种细胞分离方法，该方法使用荧光激活的细胞分选器选择低Hoescht 33342/低罗丹明123（低Ho/低Rh）荧光细胞组分，这些荧光细胞组分高度富集于LTR-HSC，并且可以重建少于20个细胞的致命辐照受体。干细胞因子(SCF) +白细胞介素-3 (IL-3) + IL-6 （90% ~ 100% HPP-CFC）存在下低Ho/低Rh细胞体外克隆具有高增殖潜力。单独Tpo不能诱导这些低Ho/低Rh细胞的增殖。然而，与SCF或IL-3联合，Tpo对细胞增殖具有多种协同作用。将Tpo添加到单个生长因子（SCF或IL-3或两者结合）中，低Ho/低Rh细胞的首次细胞分裂时间明显缩短，克隆效率大幅提高。此外，Tpo在培养早期显著增强了随后的克隆扩增。低Ho/低Rh细胞在分选后直接在琼脂中检测时，在任何生长条件下都没有形成巨核细胞集落。需要在多种细胞因子存在下培养数天才能获得集落形成单位-巨核细胞（CFU-Mk）。相比之下，分化程度更高的低Ho/高Rh细胞，先前显示含有短期再生的造血干细胞（STR-HSC），在分选后直接在Tpo中培养时，能够在琼脂中形成巨核细胞集落。这些数据表明，Tpo直接作用于使用Ho/Rh方法选择的原始造血干细胞，但这种作用依赖于多能细胞因子的存在，这些细胞随后分化为CFU-Mk。它们能够单独对Tpo作出反应。结合先前报道的Tpo对红系祖细胞的影响，这些结果表明Tpo对造血的影响比最初预期的要大。(C) 1996年由美国血液病学会出版。
In this study, we explored whether thrombopoietin (Tpo) has a direct in vitro effect on the proliferation and differentiation of long-term repopulating hematopoietic stem cells (LTR-HSC). We previously reported a cell separation method that uses the fluorescence-activated cell sorter selection of low Hoescht 33342/low Rhodamine 123 (low Ho/low Rh) fluorescence cell fractions that are highly enriched for LTR-HSC and can reconstitute lethally irradiated recipients with fewer than 20 cells. Low Ho/low Rh cells clone with high proliferative potential in vitro in the presence of stem cell factor (SCF) + interleukin-3 (IL-3) + IL-6 (90% to 100% HPP-CFC). Tpo alone did not induce proliferation of these low Ho/low Rh cells. However, in combination with SCF or IL-3, Tpo had several synergistic effects on cell proliferation. When Tpo was added to single growth factors (either SCF or IL-3 or the combination of both), the time required for the first cell division of low Ho/low Rh cells was significantly shortened and their cloning efficiency increased substantially. Moreover, the subsequent clonal expansion at the early time points of culture was significantly augmented by Tpo. Low Ho/low Rh cells, when assayed in agar directly after sorting, did not form megakaryocyte colonies in any growth condition tested. Several days of culture in the presence of multiple cytokines were required to obtain colony-forming units-megakaryocyte (CFU-Mk). In contrast, more differentiated, low Ho/high Rh cells, previously shown to contain short-term repopulating hematopoietic stem cells (STR-HSC), were able to form megakaryocyte colonies in agar when cultured in Tpo alone directly after sorting, These data establish that Tpo acts directly on primitive hematopoietic stem cells selected using the Ho/Rh method, but this effect is dependent on the presence of pluripotent cytokines, These cells subsequently differentiate into CFU-Mk, which are capable of responding to Tpo alone. Together with the results of previous reports of its effects on erythroid progenitors, these results suggest that the effects of Tpo on hematopoiesis are greater than initially anticipated. (C) 1996 by The American Society of Hematology.