伊氏锥虫体外培养株的建立及其HGPRT活性测定  

Establishment of Trypanosoma evansi in vitro Cultivation Strain and Determination of Its HGPRT Activities

作　　者：王祥生[1,2] 孙恩贵 刘小芳 杨发青[1,2] 谢超 董文其[1,2] 顾为望 

机构地区：[1]解放军农牧大学军事兽医研究所 [2]第一军医大学热带医学研究所

出　　处：《中国兽医学报》1997年第1期39-44,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金

摘　　要：用带虫培养法和带虫传代法建立了伊氏锥虫体外培养株，无论有、无饲养细胞，虫体均能快速增殖。培养７０ｄ以上，虫体仍保持原有生物学特性，对小鼠有感染性，活力旺盛，在体外能连续培养传代。虫数最高达２．４８×１０６／ｍＬ，群体增倍时间为８．８６～１０．８ｈ。液氮保存、复苏后的虫体可在饲养细胞上立即增殖。培养中发现由巨噬细胞转化的巨核母细胞对虫体生长有促进作用，经胰酶消化能与虫体一道连续传代。通过ＨＡＴ选择性培养液测定，伊氏锥虫对氨基喋呤不敏感，试验组虫体与对照一样生长良好，证明伊氏锥虫具有次黄嘌呤鸟嘌呤磷酸核糖转化酶（ＨＧＰＲＴ）A Trypanosoma evansi in vitro cultivation strain had been established by both techniques of in vitro cultivation and passage with the parasite and macrophagus in peritoneal liquid of infected mice. The parasite strain could grow and proliferate rapidly whether there was a feeder layer of macrophagus or not. T.evansi strain maintained their infectivity to mice, high vitality after being cultured and passaged in vitro continuously for 70 days. The most number of the parasite had reached 2.48 ×10 6/mL with population doubling time (P DT ) being 8.86 to 10.8 hours. The parasite strain could proliferate immediately on the feeder layer even after being preserved by liquid nitrogen. The results showed that the macronuclear blast cell transformed from macrophagus may support the parasite to grow and passage. T.evansi neither in blood of infected mice nor in vitro cultivation appeared to be sensitive to aminopurine by determination in HAT selective medium proving that there are some hypoxanthine guanine phosphoribosyltransferase (HGPRT) activities in T. evansi.

关 键 词：伊氏锥虫 体外培养 巨核母细胞 HGPRT 

分 类 号：S852.72[农业科学—基础兽医学]