The first steps towards generating induced pluripotent stem cells from cryopreserved skin biopsies of marine mammals

Abstract

A new approach for restoring populations of endangered marine mammal species is to use induced pluripotent stem cells (iPSCs). These cells can be obtained by genetic reprogramming of somatic animal cells, such as skin fibroblasts. From there, the iPSCs can be differentiated into all cell types of the organism, including spermatozoa and oocytes. The skin biopsies of three species of marine mammals, belonging to different orders (the walrus Odobenus rosmarus, the Steller sea lion Eumetopias jubatus and the Irrawaddy dolphin Orcaella brevirostris), were taken and cryopreserved. The fibroblast cultures were obtained from thawed skin biopsies of sea lion and walrus using previously developed methods for human cells. In contrast, Irrawaddy dolphin skin cells did not survive this cell isolation procedure. We had to modify the isolation method to obtain a fibroblast culture from this animal, using mechanical disaggregation only and a higher concentration of serum and antibiotics at the first steps of cultivation. The functionally active fibroblasts will be used in the next step, generating iPSCs. Testing the quality of the fibroblast cultures, we found that the abnormal fibroblasts differed in size and the distribution of filamentous actin and tubulin, compared to normal cells, and demonstrated clear aberrations in the nuclei.