人胎肺细胞培养液中类血小板生成素的初步研究

INITIAL RESEACH OF THROMBOPOIETIN LIKE PROTEIN IN CONDITIONED CULTURE MEDIUM OF HUMAN FETAL LUNG CELLS

人正常胎肺细胞用含１０％小牛血清的ＭＥＭ培养液原代培养，后用无血清的１９９培养液继续培养．硝酸纤维膜的斑点酶联免疫印迹法测定显示所收集的无血清培养液中含有能与人重组血小板生成素单抗结合的蛋白质．免疫细胞化学检测发现培养的肺细胞与血小板生成素单抗有明显阳性反应．用小鼠骨髓乙酰胆碱酯酶阳性细胞法测定肺细胞在不同培养时间所收集的培养液促血小板生成活性的大小，结果显示收集的培养液有明显刺激活性并且其活性与肺细胞的密度有相关性．

Normal diploid cells prepared from human fetal lung tissue were cultured in MEM medium contained 10% neo natal calf serum, then the cells were cultured in the serum free 199 medium. The collected serum free culture medium was proved containing a thrombopoietin (TPO) like protein which could bind to the human recombination TPO monoclonal antibodies, using nutrocellulose membrane dot ELISA. Immunohistochemistry analysis was shown that the cultured lung cells had obvious positive reaction with TPO monoclonal antibodies. A staining procedure for acetylcholinestersae with the mice femurs marrow cells after serum free culture was used to determine the thrombopoiesis activity of the culture medium collected from different cultured time's lung cells. It was demonstrated that the medium could remarkably stimulate thrombopoiesis and the activity was related with the density of the cultured lung cells.