Hill R J, Leven R M, Levin F C, Levin J
Department of Laboratory Medicine, University of California School of Medicine, San Francisco.
Exp Hematol. 1989 Sep;17(8):903-7.
Enriched populations of guinea pig bone marrow megakaryocytes were prepared by density gradient and velocity centrifugation and maintained in liquid cultures for 24 or 48 h. The resulting megakaryocyte preparations were of 86.1% +/- 5.5% purity. After 24 or 48 h in liquid culture, recovery of viable cells was 77% +/- 11% or 83% +/- 13%, respectively. Megakaryocyte cultures were supplemented with 100-200 micrograms/ml of either a lectin-fractionated preparation of thrombopoietin (TPO) from the plasma of thrombocytopenic rabbits or an identically prepared protein fraction from non-thrombocytopenic animals. Addition of TPO resulted in a significant increase (p less than 0.05) in both the proportion and total numbers of 32N megakaryocytes and a significant decrease (p less than 0.05) in the relative frequency of 8N megakaryocytes. In most experiments, a decrease in the total number of 8N megakaryocytes also was noted. These results indicate that partially purified TPO is able to increase the ploidy (DNA levels) of megakaryocytes in vitro.
通过密度梯度和速度离心制备富含豚鼠骨髓巨核细胞的群体，并在液体培养中维持24或48小时。所得巨核细胞制剂的纯度为86.1%±5.5%。在液体培养24或48小时后，活细胞的回收率分别为77%±11%或83%±13%。巨核细胞培养物补充有100 - 200微克/毫升的以下两种物质之一：来自血小板减少兔血浆的血小板生成素（TPO）的凝集素分级分离制剂，或来自非血小板减少动物的相同制备的蛋白质级分。添加TPO导致32N巨核细胞的比例和总数均显著增加（p < 0.05），而8N巨核细胞的相对频率显著降低（p < 0.05）。在大多数实验中，还观察到8N巨核细胞总数减少。这些结果表明，部分纯化的TPO能够在体外增加巨核细胞的倍性（DNA水平）。
