伊氏锥虫体外培养的研究——Ⅰ.培养系统的建立

STUDIES ON CULTIVATION OF TRYPANOSOMA EVANSI IN VITRO——Ⅰ.ESTABLISHMENT OF CULTIVATION SYSTEM

以199培养基添加25.0mmol/L Hepes,5.5mmol/L D 葡萄糖,0.2mmol/L 2-巯基乙醇,2.0mmol/L 丙酮酸钠和2.0mmol/L L 谷氨酰胺为培养液,并加20%的灭活马血清,分别以牛肺成纤维细胞和小牛胸腺成纤维细胞为饲养层细胞组成两种组合;或以同样的培养液改加20%灭活犊牛血清或20%灭活驴血清,以牛肺成纤维细胞作为饲养层细胞组成另两种组合。用这4种组合在37℃和烛缸的气相条件下成功地连续培养伊氏锥虫江苏高邮株60d 以上,培养的虫体对小白鼠的感染性和致病力无明显变化。

JG strain of T.evansi was successfully cultured in medium 199,supplemented with 25.0mmol/L HEPES,5.5 mmol/L D-glucose,0.2 mmol/L 2-ME,2.0 mmol/L Na-pyruvate,2.0mmol/L L-glutamine,10% or 20% heat-inactivated horse serum at 37℃ in the presence of bovine lungfibroblast or bovine thymus fibroblast and the same medium,20% heat-inactivated calf serum or 20%heat-inactivated donkey serum,at 37℃ in the presence of bovine lung fibroblast.All cultivation was madein a candel jar.——those combinations could support the growth of T.evansi JG strain in vitro for more than60 days.The cultured trypanosomes of T.evansi JG retained their infectivity and pathogenicity to mice.