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.
Biobanking of viable cells is essential to provide long-term security of wildlife existence. Current cell technology enables us to cultivate primary cells and adult germ cells from tissues of living and postmortem fishing cats for cryopreservation. The primary cells exhibited limited proliferation and cell senescence, which can be overcome by reprogramming the somatic cells toward pluripotent state. Here we explored the challenge of tissue collection from fishing cat and several virus-free approaches to induce cellular reprogramming in the fishing cat cells and provided insight into the techniques and conditions to enhance cell expansion, which support the success of generation of fishing cat cell biobank.