Purification of maturation-induced factor of megakaryoblastic cell line

从白血病细胞系上清液、再生障碍性贫血患者尿液和兔血小板减少性血浆中提纯了一种诱导巨核细胞系成熟的体液因子。用该材料研究了(1)小鼠巨核细胞系L8057乙酰胆碱酯酶(AchE)的表达，(2)人巨核细胞系CMK糖蛋白IIb/IIIa的表达，(3)雷公藤诱导CMK细胞进入S期的生物学效应。(4)小鼠骨髓细胞AchE的表达。在白血病细胞株的培养上清中未发现该活性，而在再生障碍性贫血患者和兔血小板减少性血浆中的尿液中未发现该活性。因此，我根据小鼠骨髓细胞AChE的表达，从再生障碍性贫血患者的尿液或兔血小板减少性血浆中提纯了该因子。由于体积较小，该因子尚未得到纯化。用硫酸铵沉淀、WGL琼脂糖凝胶、蓝色琼脂糖凝胶、镍离子琼脂糖凝胶和凝胶过滤6步从兔血小板减少性血浆中分离纯化蛋白。有趣的是，这种蛋白质能刺激成熟的巨核细胞形成原血小板。

A humoral factor which induces the maturation of megakaryoblastic cell line was purified from the sources including the supernatant of leukemic cell lines, the urine obtained from the patients with aplastic anemia and rabbit thrombocytopenic plasma. Biological effects such as (1) Achetylcholinesterase (AchE) expression of murine megakaryoblastic cell line L8057, (2) Glycoprotein IIb/IIIa expression of human megakaryoblastic cell line CMK (3) The raito to induce CMK cells to S-phase (4) AchE expression of murine bone marrow cells were studied using this materials. The activity was not found in the supernatant of leukemic cell lines, but in the urine obtained from the patients with aplastic anemia and rabbit thrombocytopenic plasma. Therefore I purified the factor from the urine obtained from the patients with aplastic anemia or rabbit thrombocytopenic plasma based on the AchE expression of murine bone marrow cells. The factor has not been purified because of small amount of volume. The protein was purified from rabbit thrombocytopenic plasma using 6 steps including ammonium sulfate precipitation, WGL sepharose, Blue sepharose, Ni^<2+> sepharose and gel filtration. Interestingly, this protein stimulated proplatelet formation from mature megakaryocytes.