Fishing Cat Cell Biobanking for Conservation

Establishment of biobank to keep wildlife cells secure long-term conservation. Fishing cat (Prionailurus viverrinus) is one of Vulnerable wild felids, currently under threaten by wetland destruction and other human activities. Here we aimed to generate cell biobanking of fishing cats by deriving various sources of primary cells from the living and postmortem animals and enhancing their expandable potency by virus-free cellular reprogramming. We show that cells can be propagated from several tissues harvested from both living and dead fishing cats with different derivation efficiency. Testes from the postmortem animals contain several tissues that can be derived primary cells as well as putative alkaline phosphatase positive and SOX2 positive adult spermatogonial stem cells. Primary cells from ear pinna and abdomen sources can only be obtained from the living fishing cats. These primary cells exhibited sign of cell senescence after a few sub-cultures, limited its usability for downstream applications. This obstacle can be overcome by reprogramming via either nucleofection or liposome-based DNA/RNA delivery. The putative iPSC colonies as well as expandable induced cells from episomal-based reprogramming appeared to be a suitable choice for expansion of cells for cryopreservation. Thus, here we provide current conservation plan using cell technology for fishing cats and also recommendation of tissue collection and culture procedures for zoo researches to facilitate the preservation of cells from postmortem animals and living animals.