Comparison of Echinostoma caproni mother sporocyst development in vivo and in vitro using Biomphalaria glabrata snails and a B. glabrata embryonic cell line

Abstract

Biomphalaria glabrata embryonic (Bge) cells have previously been shown to permit a successful cocultivation of Schistosoma mansoni and Schistosoma japonicum from miracidia to mother sporocysts (MS) and then to the production of daughter sporocysts (DS). To investigate further the properties of the Bge culturing system we used Echinostoma caproni under identical in vitro conditions. In vitro-derived miracidia were used either for experimental infections of B. glabrata snails, or for in vitro cultivation with Bge cells. Histological analysis showed that the development of MS in B. glabrata was similar to the previously described development in Biomphalaria pfeifferi in terms of final site of infection, development dynamics, growth dynamics, reproduction intensity, and life spans. Only short delays in migration dynamics were observed in B. glabrata. When cultivated under in vitro conditions, E. caproni MS could live for up to 17 wk in the presence of Bge cells, as compared with 2 wk in cell-free Bge medium. The presence of Bge cells also permitted significant growth of MS and development through complete embryogenesis of the next intramolluscan stage (embryos of 100-110 cells). However, degeneration of MS consistently occurred before production of this second generation. During the entire cultivation period, no visible contact was observed between MS and Bge cells, suggesting that development of MS was only triggered by soluble factors released by Bge cells.