微细胞(microcell)的制备及其生物学特性

MICROCELLS,THEIR PREPARATION AND BIOLOGICAL CHARACTERISTICS

本文比较了单纯用秋水酰胺(1μg/ml)和秋水酰胺(1μg/ml)加细胞松弛素B(1μg/ml)复合处理体外培养的细胞株的微细胞生产率,发现后一种方法大大提高了微细胞的获得率。对于微核化细胞的离心脱核方法以及粗制的微细胞纯化方法亦作了改进。同时研究了微细胞的存活时间以及它的大小等生物学特性。并且研究了CHO Wg3-h SL7 HFC四种细胞在秋水仙胺和秋水仙胺加细胞松弛素B复合处理后的微核率,发现后一种方法能促进细胞的微核化,但不同的细胞株对这两种有丝分裂阻断剂敏感性差异很大,体外未经转化的人的正常成纤维细胞对有丝分裂阻断剂的敏感性与细胞年龄有密切关系,越年轻越敏感。

1.The exponentially growing population of CHO cells(Chinese hamster ovarian fibroblast)and Wg3-h cells(Chinese hamster lung fibroblast)were micronucleated by treatments in succession with colcemid(1μg/ml,24hr)and with cytochalasin B(5μg/ml,12hr).The micronucleated cells attached to the plastic slides were enucleated by centrifugation at 19,000 r.p.m.for 30 min in complete medium plus cytochalasin B(10μg/ml)at 34-37°C.The plastic slides were cut from tissue culture flasks with heating needle.The crude microcell preparations were purified by filtration through microporous membrane filters.The survival rates and the diameters of the microcells were studied.In addition,the yield of microcells was counted in each experiment.2.The sensitivities of CHO,Wg3-h,SL7,(Human embryo lung fibroblast),and HFC(Human foreskin fibroblast)cells to mitotic arrest agents were compared.The micronucleation rates of the four cell lines produced by colcemid and cytochalasin B tretments in succession were much higher than that induced simply by colcemid.