Cicuttini F M, Welch K L, Boyd A W
Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia.
Exp Hematol. 1994 Dec;22(13):1244-51.
In this study, we show how Rhodamine-123 (Rh-123), as in other hematopoietic populations, can be used to define functionally distinct progenitor cells from human umbilical cord blood (HUCB). CD34+ cells were subdivided into Rh-123high (78.2 +/- 4.5%) and Rh-123low (21.8 +/- 3.6%). While 9.3 +/- 1.6% of the CD34+Rh-123high cells formed colonies in agar, only 0.4 +/- 0.2% of the CD34+Rh-123low population did so. However, the CD34+Rh-123low cells resulted in the greatest expansion of colony-forming cells (CFC) when cultured in liquid medium with different cytokine combinations. When the CD34+Rh-123low cells were cultured for 7 days with stem cell factor (SCF) and erythropoietin (Epo), the CD34+Rh-123low cells resulted in a 94-fold increase in CFC compared with a 2.5-fold increase from the CD34+Rh-123high cells. The combination of SCF and Epo or granulocyte-macrophage colony-stimulating factor (GM-CSF) supported the production and maintenance of CFC from CD34+Rh-123low cells > 28 days compared with only 21 days for the CD34+Rh-123high cells. Coculture of CD34+Rh-123low cells with stromal cell line 11 (SCL11) demonstrated that long-term culture initiating cells (LTCIC) were present within this population, as CFC could be recovered for > 10 weeks compared with < 6 weeks in cocultures with CD34+Rh-123high cells. The duration of maintenance of CFC in liquid culture could be further enhanced by the addition of an antibody (Ab) directed against the binding site of the GM-CSF receptor. The addition of anti-GM-CSF receptor Ab to cultures of CD34+Rh-123high and low cells supplemented with SCF, interleukin-3 (IL-3), and IL-6 resulted in an initial 10-fold decrease in CFC in cultures of both the CD34+Rh-123high and low cells. Although very few CFCs were present by 42 days in liquid cultures of CD34+Rh-123high cells, the number of CFCs in these cultures was significantly increased when anti-GM-CSF receptor Ab was added. Although this effect was also observed in cultures of CD34+Rh-123low cells, it was less dramatic as more CFC persisted even in the absence of Ab. The possible mechanism of this effect is discussed.
在本研究中，我们展示了若丹明 - 123（Rh - 123）如何像在其他造血细胞群体中一样，用于从人脐带血（HUCB）中定义功能不同的祖细胞。CD34 + 细胞被细分为Rh - 123高表达（78.2 +/- 4.5%）和Rh - 123低表达（21.8 +/- 3.6%）群体。虽然9.3 +/- 1.6%的CD34 + Rh - 123高表达细胞在琼脂中形成集落，但CD34 + Rh - 123低表达群体中只有0.4 +/- 0.2%能形成集落。然而，当在含有不同细胞因子组合的液体培养基中培养时，CD34 + Rh - 123低表达细胞导致集落形成细胞（CFC）的扩增最为显著。当CD34 + Rh - 123低表达细胞与干细胞因子（SCF）和促红细胞生成素（Epo）一起培养7天时，与CD34 + Rh - 123高表达细胞2.5倍的增加相比，CD34 + Rh - 123低表达细胞导致CFC增加了94倍。SCF与Epo或粒细胞 - 巨噬细胞集落刺激因子（GM - CSF）的组合支持CD34 + Rh - 123低表达细胞产生和维持CFC超过28天，而CD34 + Rh -
