Povolny B T, Lee M Y
Department of Oral Biology, University of Washington Schools of Dentistry and Medicine, Seattle 98195.
Exp Hematol. 1993 Apr;21(4):532-7.
Evidence suggests that among the myeloid colony-stimulating factors (CSFs), macrophage-CSF (M-CSF) is important in normal development of osteoclasts. To understand the role of M-CSF and other myeloid CSFs on clonal growth of osteoclast progenitors, isolated monkey bone marrow cells expressing the CD34 antigen were grown in soft agar or methylcellulose cultures in the presence of recombinant human M-CSF, granulocyte-macrophage CSF (GM-CSF), interleukin-3 (IL-3), calcitriol and various combinations of these factors. Day-10 to day-20 colonies were stained for tartrate-resistant acid phosphatase (TRAP) in order to identify putative osteoclast progenitors. Colonies were scored as follows: if greater than 80% of the cells in a colony contained TRAP it was scored as TRAP-positive; between 20% and 80%, TRAP-mix; and less than 20%, TRAP-negative. Further identification of osteoclast progenitors was made by simultaneous autoradiographic detection of calcitonin receptors on TRAP-stained colonies grown in methylcellulose. Among the 3 CSFs tested alone, GM-CSF stimulated the greatest number of TRAP-positive colonies; however, M-CSF and IL-3 in combination stimulated an even greater number of TRAP-positive colonies at all time points. Such additive effect was not observed between GM-CSF and IL-3. When calcitriol was added, the following increases in day-20 TRAP-positive colonies occurred: with M-CSF, 189%; with IL-3, 109%; with the combination of M-CSF and IL-3, 52%; and with GM-CSF there was no change. In the presence of calcitriol, 75% of IL-3 plus M-CSF-induced TRAP-containing colonies demonstrated calcitonin receptors over their TRAP-positive cells, whereas with M-CSF, GM-CSF or IL-3 each alone, 0 to 25% of TRAP-containing colonies had calcitonin receptor over TRAP-positive cells. These results suggest that IL-3, M-CSF and calcitriol stimulate a subpopulation of hemopoietic progenitors, likely osteoclast progenitors, that can give rise to TRAP-positive cells simultaneously expressing calcitonin receptors.
有证据表明，在髓系集落刺激因子（CSF）中，巨噬细胞集落刺激因子（M-CSF）对破骨细胞的正常发育很重要。为了解M-CSF和其他髓系CSF对破骨细胞祖细胞克隆生长的作用，将表达CD34抗原的分离猴骨髓细胞在重组人M-CSF、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素-3（IL-3）、骨化三醇以及这些因子的各种组合存在的情况下，在软琼脂或甲基纤维素培养基中培养。对第10天至第20天的集落进行抗酒石酸酸性磷酸酶（TRAP）染色，以鉴定假定的破骨细胞祖细胞。集落评分如下：如果集落中超过80%的细胞含有TRAP，则评为TRAP阳性；在20%至80%之间，评为TRAP混合；低于20%，评为TRAP阴性。通过对在甲基纤维素中生长的TRAP染色集落上降钙素受体的同步放射自显影检测，进一步鉴定破骨细胞祖细胞。在单独测试的3种CSF中，GM-CSF刺激产生的TRAP阳性集落数量最多；然而，M-CSF和IL-3联合在所有时间点刺激产生的TRAP阳性集落数量更多。GM-CSF和IL-3之间未观察到这种相加效应。加入骨化三醇后，第20天TRAP阳性集落增加如下：M-CSF组增加189%；IL-3组增加109%；M-CSF和IL-3联合组增加52%；GM-CSF组无变化。在骨化三醇存在的情况下，75%的IL-3加M-CSF诱导的含TRAP集落在其TRAP阳性细胞上显示有降钙素受体，而单独使用M-CSF、GM-CSF或IL-3时，0至25%的含TRAP集落在TRAP阳性细胞上有降钙素受体。这些结果表明，IL-3、M-CSF和骨化三醇刺激造血祖细胞亚群，可能是破骨细胞祖细胞，它们可产生同时表达降钙素受体的TRAP阳性细胞。
