Morphological, Ultrastructural, and Immunocytochemical Characterization and Assessment of Puma (Puma concolor Linnaeus, 1771) Cell Lines After Extended Culture and Cryopreservation
- 1 December 2022
- journal article
- research article
- Published by Mary Ann Liebert Inc in Biopreservation and Biobanking
- Vol. 20 (6), 557-566
- https://doi.org/10.1089/bio.2021.0117
Abstract
Cell lines are valuable tools to safeguard genetic material from species threatened with extinction that is mainly due to human action. In this scenario, the puma constitutes a species whose population is being rapidly reduced in the ecosystems it inhabits. For the first time, we characterized puma skin-derived cell lines and assessed these cells after extended culture (experiment 1) and cryopreservation (experiment 2). Initially, we identified and characterized four dermal fibroblast lines using morphology, ultrastructure, and immunofluorescence assays. Moreover, we evaluated the effects of culture time (1st, 3rd, and 10th passages) and cryopreservation on their morphology, ultrastructure, viability, metabolism, proliferative activity, reactive oxygen species (ROS) levels, mitochondrial membrane potential (ΔΨm), and apoptosis. The cells showed a typical spindle-shaped morphology with centrally located oval nuclei. The cells were identified as fibroblasts by staining for vimentin. In vitro culture after the 1st, 3rd, and 10th passages did not alter most of the evaluated parameters. Cells in the 3rd and 10th passages showed a reduction in ROS levels (p < 0.05). The ultrastructure revealed morphological damage in the prolongments, and nuclei of cells derived from the 3rd and 10th passages. Moreover, cryopreservation resulted in a reduction in ΔΨm compared with that of noncryopreserved cells, suggesting that the optimization of cryopreservation methods for puma fibroblasts is essential. In conclusion, we found that viable fibroblasts could be obtained from puma skin, with slight changes after the 10th passage in in vitro culture and cryopreservation. This is the first report on the development of cell lines derived from pumas.Keywords
This publication has 41 references indexed in Scilit:
- Comparação entre duas concentrações de glicerol para a criopreservação de sêmen de suçuarana (Puma concolor)Pesquisa Veterinária Brasileira, 2013
- Cell Cycle Synchronization of Skin Fibroblast Cells in Four Species of Family FelidaeReproduction in Domestic Animals, 2012
- Influence of cell culture configuration on the post-cryopreservation viability of primary rat hepatocytesBiomaterials, 2012
- Inducing pluripotency in somatic cells from the snow leopard (Panthera uncia), an endangered felidTheriogenology, 2012
- Cryopreservation of human bone marrow‐derived mesenchymal stem cells with reduced dimethylsulfoxide and well‐defined freezing solutionsBiotechnology Progress, 2010
- Establishment and characterization of a fibroblast cell line derived from Jining Black Grey goat for genetic conservationSmall Ruminant Research, 2009
- Developing biological resource banks as a supporting tool for wildlife reproduction and conservation: The Iberian lynx bank as a model for other endangered speciesAnimal Reproduction Science, 2009
- Cell cycle analysis and interspecies nuclear transfer of in vitro cultured skin fibroblasts of the Siberian tiger (Panthera tigris Altaica)Molecular Reproduction and Development, 2006
- In Vitro Development of Marbled Cat Embryos Derived from Interspecies Somatic Cell Nuclear TransferReproduction in Domestic Animals, 2006
- Oocyte recovery, maturation and fertilization in vitro in the puma (Felis concolor)Reproduction, 1990