The In vitro Cultivation of Nippostrongylus muris to the Adult Stage

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Abstract
Nippostrongylus muris was successfully cultured in cell-free media under axenic conditions in vitro through the free-living and parasitic cycles. Development comparable to advanced lung stages in the rat has been obtained in 50% chick embryo extract alone. Further growth and differentiation, comparable to 4th- and 5th-stage worms in the gut, occurred only when embryo extract was supplemented in a variety of ways. Fifth-stage worms were obtained in media combining the following components sodium caseinate or casein hydrolysate, filtrate of powdered yeast or liver extract, chick embryo extract, and mammalian serum. Sexually mature males containing sperm and females containing eggs have appeared so far only in a medium containing caseinate, liver filtrate, chick embryo extract and rat serum. The adult worms were morphologically normal, although small in size; the males ranged from 1.8 to 2.4 mm, and the females from 2.1 to 2.5 mm. It has taken at least 3 to 4 times longer for 5th-stage worms to develop in culture, than in the rat. Mating apparently has not occurred, due probably to the small numbers of adult worms which have appeared at any one time. Test tube cultures have proved to be considerably more advantageous than flask cultures. They are easier to handle, and allow continued observation of the organisms in the media.