Original Article

Marine Biotechnology

, Volume 15, Issue 3, pp 357-367

First online:

Primmorphs Cryopreservation: A New Method for Long-Time Storage of Sponge Cells

  • Francesca MussinoAffiliated withDIMES, University of Genoa
  • , Marina PozzoliniAffiliated withDISTAV, University of Genova
  • , Laura ValisanoAffiliated withDISTAV, University of Genova
  • , Carlo CerranoAffiliated withDISVA, Università Politecnica delle Marche
  • , Umberto BenattiAffiliated withDIMES, University of GenoaCenter of Excellence for Biomedical Research, University of Genoa
  • , Marco GiovineAffiliated withDISTAV, University of GenovaCenter of Excellence for Biomedical Research, University of GenoaDipartimento di Scienze della Terra, dell’Ambiente e della Vita (DISTAV), Università degli Studi di Genova Email author 

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The possibility to cryopreserve cells allows for wide opportunities of flexible handling of cell cultures from different sponge species. Primmorphs model, a multicellular 3D aggregate formed by dissociated sponge cells, is considered one of the best approaches to establish sponge cell culture but, in spite of the available protocols for freezing sponge cells, there is no information regarding the ability of the latter to form primmorphs after thawing. In the present work, we demonstrate that, after a freezing and thawing cycle using dissociated Petrosia ficiformis cells as a model, cells viability was high but it was not possible to obtain primmorphs. The same protocol for cryopreservation was then used to directly freeze primmorphs. In this second case, after thawing, viability and the cellular proliferative level were similar to unfrozen standard primmorphs. Spiculogenesis in thawed primmorphs was evaluated by quantifying the silicatein gene expression level and by assaying the silica amount in the newly formed spicules, then compared with the correspondent values obtained in standard unfrozen primmorphs. Results indicate that the freezing cycle does not affect the spiculogenesis rate. Finally, the expression level of heat shock protein 70, a well-known stress marker, was assayed and the results showed no differences between frozen and unfrozen samples. These findings are likely to promote relevant improvements in sponge cell culture technique, allowing for a worldwide exchange of living biological material, paving the way for cell banking of Porifera.


Porifera Cryopreservation Primmorphs