Formation of spicules by sclerocytes from the freshwater spongeEphydatia muelleri in short-term cultures in vitro

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Abstract
Cells from the freshwater spongeEphydatia muelleri were isolated by dissociating hatching gemmules. During the first 24 h the cells reaggregated, but the aggregates progressively disintegrated again to single cells, among which the spicule-forming sclerocytes were recognized. Such cultures were used to study spicule (megascleres) formation in vitro. The isolated sclerocytes formed the organic central axial filament onto which they deposited inorganic silicon. The size of the spicules (200 to 350µm in length) as well as the rate of spicule formation (1 to 10µm/h) under in vitro conditions were similar to the values measured in vivo. Immediately after completion of spicule formation, or even before, the sclerocyte could start formation of a new spicule; 5% of the cells were in the process of forming two spicules simultaneously. Cultivation of sclerocytes in the absence of silicon resulted in the formation of the axial filament only. We succeeded in maintaining the sclerocytes in a proliferating and spicule-forming state for up to 3 mo. These results demonstrate that the establishment of short-term cell cultures fromE. muelleri is possible; however, future studies must be undertaken to identify the growth factors required for a permanent culture of sponge cells.