Townson S, Connelly C, Muller R
J Helminthol. 1986 Dec;60(4):323-30. doi: 10.1017/s0022149x00008579.
A series of experiments examined the effects of various media, serum supplements, gas phases and the incorporation of mammalian cell feeder layers on the survival of Onchocerca gutturosa adult worms in vitro. The survival of male worms was poor in all media tested that were not supplemented with inactivated foetal calf serum (IFCS), with improved but variable survival in media supplemented with 10-30% IFCS. Using a cell-free system in an atmosphere of 5% CO2 in air, good results were obtained in medium NCTC 135 + 10% IFCS (median survival time 39 days, range 25-41). Marginally better survival was obtained with the same medium in an atmosphere of 95% N2/5% CO2 (median 45 days, range 25-56) and with a 1:1 mixture of media NCTC 135 and IMDM + 10% IFCS (median 38 days, range 38-51). Survival was enhanced in culture systems which incorporated bovine kidney (MDBK) cells, bovine trachea (EBTR) cells and monkey kidney (LLCMK2) cells. Exceptionally long survival was obtained using medium MEM + 10% IFCS + LLCMK2 cells under a gas phase of 5% CO2 in air, in which male worms survived from approximately 6 to over 7 months. Under similar conditions, female worms were also maintained for periods of up to 6 months and 5 out of 18 specimens released microfilariae into the culture system. The long-term culture described in this study will be useful for basic biochemical, chemotherapeutic and immunological studies in vitro.
一系列实验研究了不同培养基、血清补充剂、气相条件以及哺乳动物细胞饲养层的加入对体外培养的喉瘤盘尾丝虫成虫存活的影响。在所有未添加灭活胎牛血清（IFCS）的测试培养基中，雄虫的存活率都很低，在添加了10%-30%IFCS的培养基中，存活率有所提高但不稳定。在含5%二氧化碳的空气环境下的无细胞体系中，使用培养基NCTC 135 + 10%IFCS取得了良好效果（中位存活时间39天，范围25-41天）。在95%氮气/5%二氧化碳的环境下，使用相同培养基存活率略高（中位45天，范围25-56天），使用培养基NCTC 135和IMDM按1:1混合并添加10%IFCS时存活率也较好（中位38天，范围38-51天）。在含有牛肾（MDBK）细胞、牛气管（EBTR）细胞和猴肾（LLCMK2）细胞的培养体系中，存活率提高。在含5%二氧化碳的空气气相条件下，使用培养基MEM + 10%IFCS + LLCMK2细胞，雄虫存活时间极长，约6至7个多月。在类似条件下，雌虫也能维持长达6个月的存活，18个样本中有5个在培养体系中释放出微丝蚴。本研究中描述的长期培养方法将有助于体外进行基础生化、化疗和免疫学研究。
