Sponges have been found to be a diverse and prolific source of compounds of
potential therapeutic value. However, the yield of bioactive compounds from
sponges is typically low and the direct harvesting of sponges to generate
sufficient quantities of bioactive materials is not feasible. The development of
techniques for the in vitro production of these compounds would thus be a
major advance. Our preliminary studies employ attachment-dependent mammalian
cell culture techniques to develop sponge cell cultures. We have shown that the
cells of the sponge Microciona prolifera are capable of attaching to the
commercially available microcarriers, Cytoline 1 and Cytodex 3, and exhibit a
degree of proliferation. Optimization of attachment conditions and media
requirements of the sponges is ongoing. Sponges associate
with a rich microbial community. Libraries of small subunit ribosomal DNA (SSU
rDNA)have been constructed for the identification of these microorganisms.
Phylogenetic analyses were performed to determine phylogenetic type and origin.
The 18S rDNA sequence of the sponge Axinella corrugata has been
determined for use in cell line validation and for the design of molecular
probes for the identification of sponge cells in culture. The consistent
association of a crenarchaeote with the genus Axinella was verified.
Similarities between the SSU rDNA sequences of clones obtained from genomic DNA
extracted from the sponge revealed eubacteria that would not normally be found
in oxygenic conditions. Further analysis is necessary to determine if a
consistent association of these bacteria with the sponge exists and if they
contribute to its metabolism and nutrition.
|
