Heyworth C M, Ponting I L, Dexter T M
Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK.
J Cell Sci. 1988 Oct;91 ( Pt 2):239-47. doi: 10.1242/jcs.91.2.239.
Haemopoietic cell growth factors are normally assayed using unfractionated marrow cells (NBM). However, using this population it is difficult to distinguish between direct versus indirect effects, because of the low incidence of colony forming cells (CFC) and the presence of possible accessory cells (which may themselves be acted upon by the growth factors and stimulated to produce other growth stimulatory or inhibitory molecules that influence the development of the CFC). Furthermore, NBM contain the whole spectrum of multipotent and lineage-restricted CFC and it is often difficult to determine precisely which populations are being stimulated to develop. This latter problem can be solved, in part, by using marrow from mice previously treated with 5-fluorouracil (5-FU): an agent that preferentially kills the more mature, actively cycling CFC but spares the proliferatively quiescent multipotent stem cells. Since the 5-FU-treated marrow also contains many possible accessory cells, however, it is again not clear whether or not the responses elicited by growth factors are due to direct or indirect effects upon the CFC. To circumvent this problem we have obtained a highly enriched population of multipotent stem cells (FACS-BM) that is free of accessory cells, and have compared the responses of these cells to NBM and to 5-FU-BM in the presence of a variety of growth factors.(ABSTRACT TRUNCATED AT 250 WORDS)
造血细胞生长因子通常使用未分级的骨髓细胞(NBM)进行测定。然而,使用这种细胞群体很难区分直接效应和间接效应,这是因为集落形成细胞(CFC)的发生率较低,并且存在可能的辅助细胞(这些细胞本身可能受到生长因子的作用,并被刺激产生其他影响CFC发育的生长刺激或抑制分子)。此外,NBM包含了全谱的多能和谱系受限的CFC,通常很难精确确定哪些细胞群体受到刺激而发育。后一个问题可以部分通过使用先前用5-氟尿嘧啶(5-FU)处理过的小鼠的骨髓来解决:5-FU是一种优先杀死更成熟、活跃循环的CFC但保留增殖静止的多能干细胞的药物。然而,由于经5-FU处理的骨髓也包含许多可能的辅助细胞,因此同样不清楚生长因子引发的反应是由于对CFC的直接作用还是间接作用。为了解决这个问题,我们获得了一个高度富集的无辅助细胞的多能干细胞群体(FACS-BM),并比较了这些细胞在多种生长因子存在下对NBM和5-FU-BM的反应。(摘要截短于250字)