Source DUVI
Currently, the process of cryopreservation of cells and tissues of terrestrial organisms has become a tool of outstanding utility for medical research, both human and veterinary. On the contrary, the use of this technique in the marine environment is still limited, and there are very few protocols that have been successful in cryopreserving fish and mollusc cells. In fact, the scarcity of effective methodologies for this procedure applicable to key cell types, such as germ cells or embryos, limits the development of research in marine biology. For this reason, the scientific community is working on the development of methodologies for the preservation of a greater number of organisms and/or cell types. The University of Vigo began working in this field in 2009, specifically in cutting-edge research into the cryopreservation of embryos and larvae of marine organisms, which began as part of Estefanía Paredes’ doctoral thesis. Now, this work has become a national patent that recognizes the only protocol for cryopreservation of early stages of sea urchin development with demonstrated viability and application in the assessment of marine pollution and aquaculture. This technology and its applications are the result of the work carried out in the laboratory of Professor Ricardo Beiras and under the direction of Juan Bellas, scientist at the Spanish Institute of Oceanography in Vigo.
A specific protocol designed for sea urchins
The cryopreservation procedure for sea urchin embryos (Paracentrotus lividus) is based on a protocol specifically designed for this use and which allows blastulas (sea urchin embryos) to be stored in liquid nitrogen so that they can be used outside the natural breeding season of this species. The patent has been granted to Ricardo Beiras, Juan Bellas and Estefanía Paredes. The latter, who currently works in the United States, explains that the patent was developed during her doctoral thesis, which received the award from the Royal Academy of Doctors of Spain in 2015, for which “much of the data was obtained at Ecimat”. The main novelty it presents is that “it develops a cryopreservation protocol of proven success for sea urchins, a milestone that had not been achieved previously”, explains Paredes, who adds that its possible application in ecotoxicology and aquaculture has also been developed. The researcher emphasizes that the patent application process is not simple, and she highly values the training that the University of Vigo offers on the protection of intellectual property or the patent system and the help and advice from the OTRI.
Paredes is currently completing her postdoctoral studies at the Laboratory of Theoretical and Applied Cryobiology at the University of Tennessee in Knoxville. “I came here,” she explains, “to work with Dr. Peter Mazur, one of the founding fathers of cryobiology, and to delve deeper into the more theoretical aspects of cryopreservation of species considered ‘model’ in science.” After Mazur’s death in 2015, the Galician researcher was in charge of his laboratory and the completion of the National Institute of Health (NIH) project they were working on.
Cryomar project pending
In the near future, Estefanía Paredes could return to Vigo with the Cryomar project. As explained by Antonio Villanueva, deputy director of Ecimat, the Vigo center is leading a subproject in the Assemble+ proposal submitted in March to the European call for proposals H2020-INFRAIA in collaboration with 24 other European institutions. The proposal is focused on the development of cryopreservation techniques for different groups of marine organisms and in the event of obtaining European funding “this project would represent a very important boost to the development of the cryopreservation service of Ecimat”. This is one of the main strategic bets of the Vigo marine science station, given that it is a tool with a very transversal application and with enormous potential to radically facilitate the study of marine biological systems. As Villanueva highlights, “the availability of marine biological resources is one of the main limitations for marine R&D due to the cost of collecting and/or maintaining organisms in captivity, the seasonality and the scarcity of specimens in the natural environment”. Cryopreservation is the main alternative for the ex situ storage of marine biological resources, and allows for reduced costs and storage space, while facilitating intact and functional preservation of organisms.
State funding for the Functional Preservation Service
Ecimat’s commitment to cryopreservation as a strategic line of R&D was reflected in the creation of a Functional Preservation Service, which is part of the Marine Cultures Unit, and which represents, as Antonio Villanueva emphasizes, “one of Ecimat’s main contributions to the European Marine Biological Resource Centre (EMBRC). For the implementation of this service, the research centre has just received significant support from MINECO through the call for the award of grants for infrastructure and scientific-technical equipment. This award, Villanueva highlights, will allow the acquisition of, among others, state-of-the-art cryopreservation equipment that will allow Ecimat to make a qualitative leap in the provision of the service due to the incorporation of the innovative cryopreservation technique by vitrification with ultra-rapid laser thawing that obtains very high survival rates after thawing”. In this way, the requested equipment “would make Ecimat one of the first laboratories worldwide to use this technology and the first to do so to cryopreserve marine organisms”.
