MEPS 178:205-219 (1999)  -  doi:10.3354/meps178205

Establishment of a primary cell culture from a sponge: primmorphs from Suberites domuncula

Werner E. G. Müller1,*, Matthias Wiens1, Renato Batel2, Renate Steffen1, Heinz C. Schröder1, Radovan Borojevic3, Marcio Reis Custodio1,3

1Institut für Physiologische Chemie, Abteilung Angewandte Molekularbiologie, Universität, Duesbergweg 6, D-55099 Mainz, Germany
2Center for Marine Research, 'Ruder Boskovic' Institute, 52210 Rovinj, Croatia
3Departamento de Histologia e Embriologia, Instituto de Ciêcias Biomédicas, Universidade Federal de Rio de Janeiro, CP 68021, Rio de Janeiro CEP 21941-970, Brazil

ABSTRACT: In spite of the fact that cells from the phylum Porifera (sponges) contain high levels of telomerase activity, no successful approach to cultivate sponge cells has yet been described. Telomerase is the enzyme which catalyzes the addition of new telomeres onto chromosome ends which have been lost after each round of DNA synthesis. One reason may be seen in the observation that after dissociation the cells lose their telomerase activity. In addition, no nutrients and metabolites have been identified that would stimulate sponge cells to divide. We report here the culture conditions required for the formation of multicellular aggregates from Suberites domuncula from dissociated single cells; they are termed primmorphs. These aggregates, formed in seawater supplemented with antibiotics, have a tissue-like appearance; they have been cultured for more than 5 mo. Cross sections through the primmorphs revealed an organized zonation into a distinct unicellular epithelium-like layer of pinacocytes and a central zone composed primarily of spherulous cells. After their association into primmorphs, the cells turn from the telomerase-negative state to the telomerase-positive state. Important is the finding that a major fraction of the cells in the primmorphs undergo DNA synthesis and hence have the capacity to divide. By applying the BrdU (5-bromo-2'-deoxy-uridine)-labeling and detection assay it is demonstrated that up to 33.8% of the cells in the primmorphs are labeled with BrdU after an incubation period of 12 h. It is proposed that the primmorph system described here is a powerful novel model system to study basic mechanisms of cell proliferation and cell death; it can also be used in aquaculture, for the production of bioactive compounds and as a bioindicator system.

KEY WORDS: Suberites domuncula · Sponges · Cell culture · Telomerase · Primmorphs · Senescence · Apoptosis · Aquaculture · Bioactive compounds · Bioindicator · Biomarker

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