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Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - DRYSTORE (Exploring dry storage as an alternative biobanking strategy inspired by Nature)

Teaser

Summary

The problem addressed.
Drystore address the possibility to induce a reversible drying in gametes and somatic cells by conferring them protection from the water stress through protecting agents (xeroprotectants) found in naturally desiccation tolerant organisms (anhydrobiontes). Biobanking under liquid nitrogen (LN) has satisfactory efficiency but it comes with many drawbacks (e.g. expensive, high carbon footprint, need dedicated facilities, trained personnel, continuous monitoring and guaranteed LN and power supply). Searching Nature for alternatives, long-term preservation there is done by drying rather than freezing. To date, however, mammalian cells do not survive the drying process. Even so, if somatic cells\' genetic and epigenetic load can be maintained, drying can find an important role in biobanking, a fast growing global industry. Bearing in mind the many limitations of cryostorage, countless uses and applications of biobanking face serious sustainability challenges.

Its importance for society.
Dry biobanking would benefit fields such as biodiversity conservation, blood banks, massive cell and tissue storage for research and precision medicine initiatives and would have worldwide forcefulness during conflicts and environmental catastrophes. Dry storage will make biobanking more accessible to a wider range of small and medium size enterprises and third world nations where liquid nitrogen and power supply are limited and often unreliable. Being considerably cheaper, dry biobanking will save huge societal expenditures, freeing funds for other important purposes.

The overall objectives.
In this project we aimed to elucidate the effects of drying on DNA structure and function and search for proper protection to the cells through the process, using somatic cells as a model, and combining advanced freezing and drying techniques with state-of-the-art Somatic Cell Nuclear Transfer (SCNT). The expected outcomes were definitive data on effects of drying on genomic stability and function, and possible discovery of a protocol to protect the genetic and epigenetic loads during cell drying.

Work performed

Personal training: included completion of personal carrier development plan, gaining experience in teaching and supervision of students, leading and/or participating in writing competitive national and international research grants applications, presenting the drying at conferences, and gaining experience in project and budget management.
Scientific training: I have learned to generate primary fibroblast cell culture, handle the cells, move them through passages and eventually cryopreserve them and/or prepare them for the analyses foreseen by the DRYSTORE project. Several primary cell lines are already ready and cryopreserved. I also received training in oocyte collection/aspiration from abbatoir-collected ovaries, performing oocyte maturation, activation and culture. Furthermore, I was trained in the process of preparing micro tools (holding pipettes, enucleation pipettes, and injection pipettes) and learned the process of intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT) using micromanipulator. Proficiency in this field requires extensive experience that I am in the process of gaining. I have also transferred from my skills to the Host. This included knowledge in sperm handling, cryopreservation, drying, and sperm biology, wildlife reproduction as well as supporting the team members with my knowledge of English and scientific writing. I have also introduced a number of personal contacts to the HOST, some of which were invited to give talks at international meetings organised by the HOST, some participated in joint grant applications, and with some there is on-going research collaboration.
Dissemination and communication: so far I have co-authored three papers - two as a lead author and one as the senior author. One of the papers (published in the Journal of Reproduction and Development) was selected by the editor as the cover article and a figure related to the article is on the cover of the October 2018 issue of the journal. I was invited to give talks at a number of international meetings as well as a lecture at a joint meeting of the Department of Animal Sciences and the School of Veterinary Medicine, both at the Robert H. Smith Faculty of Agriculture, Food, and Environment of the Hebrew University, Rehovot, Israel. A press conference announcing the start of the project was held in October 2017 and received wide local and national coverage. The project website was launched (https://drystore.unite.it). I was interviewed twice for Channel B of the Israeli Public Broadcasting Corporation. Links (in Hebrew) are available on DRYSTORE website.

Final results

Our aim was, and still is, to find the way to a safe, reliable, friendly, low-cost approach to establish and maintain biobanks. DRYSTORE has advanced the knowledge in the field of desiccation, thus enhancing Europe as a leading actor in non-cryogenic storage technology for biobanks. By the end of the project we have learned that freeze-drying damages the DNA on both the single- and double-strand levels but that some of the damage can be protected against by xeroprotectantys and some may be corrected by the oocytes, that electroporation is a useful tool to load the cells with xeroprotectants but membrane-permeable xeroprotectants (in development) would potentially be a better and less damaging option. We have learned that slow rehydration better maintains cell morphology and we have optimised the drying process to achieve water content of around 5%. It is a bit premature to talk about economic impact or societal implications of the project at this stage. Work continues after the end of the project. The outcome of the project would act as a leverage to apply for other competitive grants to continue the research to the point where it would achieve the technology\'s promising potential.