Farrar R G, Klei T R
J Parasitol. 1985 Aug;71(4):489-99.
Strongylus edentatus was successfully cultured in vitro to the fourth larval stage (L4). Some growth continued for periods of 40-50 days at which time reductions in viability were observed in some of the culture systems tested. Various combinations of media, sera, buffers and organ explant cultures were tested. All cultures were incubated at 37 C in an atmosphere of 95% air and 5% CO2. Larvae underwent growth and differentiation to the L4 in all medium-serum combinations with and without organ explant cultures. Development and growth did occur but viability was reduced to insignificant levels in media without serum or cells. Optimal growth, differentiation, and longevity were observed in bicarbonate buffered RPMI-1640 containing 10% fetal calf serum and gerbil (Meriones unguiculatus) cecum explant cultures. Observations indicated that Strongylus vulgaris and Strongylus equinus also developed to the L4 stage using similar techniques. However, viability of S. vulgaris L4 was markedly limited. Specific morphological changes marked phases of development of S. edentatus, categorized as early, middle and late third stage, third molt and early fourth stage. Strongylus equinus appeared to follow the same developmental pattern in vitro as S. edentatus. Distinct differences in morphological features during differentiation were observed between S. edentatus and S. vulgaris.
无齿圆线虫成功在体外培养至第四幼虫期（L4）。在40 - 50天的时间段内仍有一定生长，此时在一些测试的培养系统中观察到活力有所下降。对培养基、血清、缓冲液和器官外植体培养的各种组合进行了测试。所有培养物均在37℃、95%空气和5%二氧化碳的气氛中孵育。无论有无器官外植体培养，幼虫在所有培养基 - 血清组合中均生长并分化至L4期。发育和生长确实发生了，但在无血清或细胞的培养基中，活力降至微不足道的水平。在含有10%胎牛血清和沙鼠（长爪沙鼠）盲肠外植体培养物的碳酸氢盐缓冲RPMI - 1640中观察到最佳生长、分化和寿命。观察表明，普通圆线虫和马圆线虫也使用类似技术发育至L4期。然而，普通圆线虫L4的活力明显受限。无齿圆线虫发育阶段的特定形态变化分为早期、中期和晚期第三阶段、第三次蜕皮和早期第四阶段。马圆线虫在体外似乎遵循与无齿圆线虫相同的发育模式。在无齿圆线虫和普通圆线虫分化过程中的形态特征观察到明显差异。
