Title: iPSC Derivation to Recover Threatened Wild Species - From reviewing techniques for obtaining iPSCs to Comparing Induction Outcomes in Avian and Human Cells

Biography:

Rachel Castro Teixeira dos Santos is engaged in research in the Cellular Biology laboratory at the Institute of Biological Sciences of the Federal University of Minas Gerais through the Institutional Program of Scholarships for Technological Development and Innovation (PIBIT). She began her research career while still in high school, being awarded at four higher education institutions: Federal Institute of Education, Science and Technology of Southeastern Minas Gerais (IFET), the Federal University of Juiz de Fora (UFJF), the Federal University of Minas Gerais (UFMG), and the University of São Paulo (USP). She worked in the field of Microbiology at the Laboratory of Biotherapeutic Agents (LABio) at the Institute of Biological Sciences of UFMG through PIBIT (2021 – 2022). She was a student of the Summer Course in Human Reproduction at the Fertility Clinic in São Paulo, where she later served as a monitor (2022-2023) and worked as an intern at the CELL Clinic – Assisted Human Reproduction (2023). Currently, she is working on the research project "Reproductive Biotechnologies with iPSC Generation for the Recovery of Endangered Avian Species" in the area of Cellular Biology at the Department of Morphology at the Institute of Biological Sciences of UFMG.Iara Pastor Nogueira is currently a doctoral candidate in the Graduate Program in Cellular Biology at the Institute of Biological Sciences (ICB) of the Federal University of Minas Gerais. She holds a Master's degree in Physiology from the Graduate Program in Biological Sciences: Physiology and Pharmacology of the ICB (2020), as well as from the Department of Physiology at Western University – Ontario – Canada, through the ELAP Program (Emerging Leaders of the Americas Program – 2019). Abroad, she also participated in the Science Without Borders program, completing the final year of the Marine Biology course at Swansea University – United Kingdom. Through this program, she interned in the Veterinary Medicine sector of the Lisbon Oceanarium – Portugal. She presented two papers as the first author at the YABOUMBA congress in Paris – France.

Abstract

Induced pluripotent stem cells (iPSCs) were first generated by Yamanaka in 2006, revolutionizing research by overcoming limitations imposed by the use of embryonic stem cells. In terms of the conservation of endangered species, iPSC technology presents itself as a viable alternative for the manipulation of target genetics without compromising specimens. Although iPSCs have been successfully generated for various species, their application in nonmammalian species, particularly avian species, requires further in-depth investigation to cover the diversity of wild species at risk and their different protocol requirements. This study aims to provide an overview of the workflow for iPSC induction, comparing well-established protocols in humans and mice with the limited information available for avian species. In addition to the review, the use of lipid nanoparticles LNP4 for inducing pluripotent stem cells in avian and human cells will be discussed.Gene delivery via LNP4 is non-integrative, providing greater biosafety and better performance compared to methods using cationic lipids. It promotes increased delivery of DNA into the cell nucleus, higher cell viability, and exhibits lower immunogenicity compared to other cell transfection methods. Here, we discuss the somatic cell sources to be reprogrammed, genetic factors, delivery methods, enhancers, a brief history of achievements in avian iPSC derivation, the main approaches for iPSC characterization, laboratory results regarding transfection using LNP4 in nonmammalian origin cells (Feather follicle Cells – FFCs – derived from Gallus gallus and Anodorhynchus hyacinthinus) and human origin cells (spontaneously immortalized keratinocytes - HaCaT) and the future perspectives and challenges for the field. By examining the current protocols and state-of-the-art techniques employed in iPSC generation, we seek to contribute to the development of efficient and species-specific iPSC methodologies for at-risk avian species. The advancement of iPSC technology holds great promise for achieving in vitro germline competency and, consequently, addressing reproductive challenges in endangered species, providing valuable tools for basic research, bird genetic preservation and rescue, and the establishment of cryobanks for future conservation efforts.