Gerson S L, Cooper R A
Leuk Res. 1984;8(5):791-800. doi: 10.1016/0145-2126(84)90100-0.
Phorbol esters have two opposing effects on bone marrow granulocyte/macrophage colony forming cells (CFC-GM): they reduce the number of CFC-GM which respond to colony-stimulating activity (CSA), and they also induce the release of CSA from accessory marrow cells. We questioned whether the direct inhibitory effect of phorbol esters on CFC-GM was reversible and whether phorbol ester-treated accessory bone marrow cells could increase the in vitro recovery of cultured CFC-GM. Normal human bone marrow cells were exposed to phorbol-12,13-dibutyrate (PDB) for 5 days in liquid cultures. Total nucleated cell counts decreased in a dose-dependent fashion to 42 +/- 6% of control at 5 X 10(-8) M PDB, and CFC-GM decreased to 20 +/- 8% of control. The cultures were then washed to remove PDB and continued for up to 8 weeks. Total cell counts and CFC-GM/ml remained reduced in PDB-pretreated cultures compared to control. To determine whether this was a direct effect of PDB on nonadherent, proliferating cells or whether PDB acted indirectly by affecting adherent, accessory cells, nonadherent bone marrow cells from control and PDB-preincubated cultures were co-cultured for 8 weeks with either control or PDB-preincubated adherent bone marrow cells. Nonadherent cells preincubated with PDB had a similar growth pattern whether or not the co-cultured adherent cells had been preincubated with PDB. In contrast, nonadherent cells preincubated in control medium and co-cultured with PDB-preincubated adherent cells displayed increased numbers of nucleated cells, CFC-GM and adherent hematopoietic islands compared to control. This was associated with increased amounts of CSA present in the culture supernatants. Thus PDB causes an irreversible decrease in CFC-GM in long term marrow cultures but enhances the ability of adherent stromal cells to support normal CFC-GM growth in culture.
佛波酯对骨髓粒细胞/巨噬细胞集落形成细胞（CFC-GM）有两种相反的作用：它们减少对集落刺激活性（CSA）产生反应的CFC-GM的数量，同时还诱导辅助骨髓细胞释放CSA。我们质疑佛波酯对CFC-GM的直接抑制作用是否可逆，以及经佛波酯处理的辅助骨髓细胞是否能增加体外培养的CFC-GM的恢复率。正常人骨髓细胞在液体培养中暴露于佛波-12,13-二丁酸酯（PDB）5天。有核细胞总数呈剂量依赖性下降，在5×10⁻⁸M PDB时降至对照的42±6%，CFC-GM降至对照的20±8%。然后洗涤培养物以去除PDB，并持续培养长达8周。与对照相比，PDB预处理的培养物中的总细胞数和每毫升CFC-GM仍然减少。为了确定这是PDB对非贴壁增殖细胞的直接作用，还是PDB通过影响贴壁辅助细胞间接起作用，将来自对照和PDB预孵育培养物的非贴壁骨髓细胞与对照或PDB预孵育的贴壁骨髓细胞共培养8周。无论共培养的贴壁细胞是否用PDB预孵育，用PDB预孵育的非贴壁细胞都有相似的生长模式。相反，与用PDB预孵育的贴壁细胞共培养的、在对照培养基中预孵育的非贴壁细胞，与对照相比，其有核细胞、CFC-GM和贴壁造血岛的数量增加。这与培养上清液中存在的CSA量增加有关。因此，PDB在长期骨髓培养中导致CFC-GM不可逆地减少，但增强了贴壁基质细胞支持培养中正常CFC-GM生长的能力。
