Porter P N, Ogawa M
Blood. 1982 Jun;59(6):1207-12.
Bone marrow conditioned media (BMCM) increases burst number and the incorporation of 59Fe into heme by bursts when peripheral blood or bone marrow cells are cultured at limiting serum concentrations. Burst-promoting activity (BPA) has now been purified approximately 300-fold from this source by ion-exchange chromatography on DEAE-Sephadex and absorption chromatography on hydroxyapatite agarose gel. Marrow BPA increased burst number and hemoglobin (Hb) synthesis in a dose-dependent manner. A larger increase in Hb synthesis than in burst number was consistently observed, which was probably a consequence of the increase in the number of cells per burst that occurs in the presence of BPA. The role of BPA in culture could be distinguished from erythropoietin (Ep), since no bursts grew in the absence of Ep, whether or not BPA was present, and since it had no effect on the growth of erythroid colonies scored at day 5 of culture. Our purified fraction did not support the growth of CFU-C in culture. Activity was stable at temperatures of 70 degrees C or lower for 10 min; exposure to 80 degrees C resulted in approximately 50% loss of activity. BPA was completely inactivated by treatment at 100 degrees C for 10 min. Thus, human bone marrow cells produce a heat-sensitive factor that specifically promotes the growth of early erythroid progenitors in culture.
当外周血或骨髓细胞在极限血清浓度下培养时，骨髓条件培养基（BMCM）可增加爆式集落形成单位数量，并使爆式集落形成单位将59Fe掺入血红素中。现在，通过在DEAE-葡聚糖上进行离子交换色谱和在羟基磷灰石琼脂糖凝胶上进行吸附色谱，已从该来源中纯化出爆式集落形成单位促进活性（BPA）约300倍。骨髓BPA以剂量依赖的方式增加爆式集落形成单位数量和血红蛋白（Hb）合成。始终观察到Hb合成的增加幅度大于爆式集落形成单位数量的增加幅度，这可能是由于在BPA存在下每个爆式集落形成单位中的细胞数量增加所致。BPA在培养中的作用可与促红细胞生成素（Ep）区分开来，因为无论是否存在BPA，在没有Ep的情况下都不会有爆式集落形成单位生长，并且因为它对培养第5天计数的红系集落生长没有影响。我们纯化的组分不支持培养中的集落形成单位-粒系（CFU-C）生长。活性在70℃或更低温度下保持10分钟稳定；暴露于80℃导致约50%的活性丧失。BPA在100℃处理10分钟后完全失活。因此，人骨髓细胞产生一种热敏感因子，该因子在培养中特异性促进早期红系祖细胞的生长。
