Long-term preservation of established fibroblast lines from six‐banded armadillos (Euphractus sexcintus, Linnaeus, 1758) by extended passage and cryopreservation

• Published in: In vitro cellular & developmental biology. Animal 2024-03, Vol.60 (3), p.266-277
• Main Authors: Fernandes, Denilsa Pires, Praxedes, Érika Almeida, da Silva Viana, João Vitor, de Oliveira Santos, Maria Valéria, Silva, Alexandre Rodrigues, Freitas, Carlos Iberê Alves, Pereira, Alexsandra Fernandes
• Format: Article
• Language: English
• Subjects: Animal Genetics and Genomics; Animals; Apoptosis; Armadillos; Biomedical and Life Sciences; Cell Biology; Cell Culture; Cell Line; Cell morphology; Cryopreservation; Cytology; Developmental Biology; Explants; Fibroblasts; Freezing; Humans; In vitro; Karyotypes; Life Sciences; Membrane permeability; Membrane potential; Membranes; Metabolism; Morphology; Skin; Somatic cells; Stem Cells; Viability; Vimentin
• ISSN: 1071-2690; 1543-706X; 1543-706X
• DOI: 10.1007/s11626-024-00871-w

Establishing new somatic cell cultures has raised significant attention as an effective and convenient way to preserve genetic samples for different applications. Although many lines have been established in model animals, none derived from six-banded armadillo species is currently available. We report the successful isolation and characterization of fibroblasts from six‐banded armadillos, evaluating the cell quality after extended culture and cryopreservation. Initially, we collected ear skin from five captive adult individuals and identified fibroblast lines by morphology, karyotyping, and immunophenotyping assays. The isolated fibroblasts were evaluated after several passages (fourth, seventh, and tenth passages) and cryopreservation by slow freezing. Cell morphology, viability, metabolism, proliferative activity, mitochondrial membrane potential, and apoptosis levels were analyzed. The skin explants had great adhesion, and cell outgrowth could be seen after 3–6 d. The cells were verified as fibroblasts at the fourth passage by vimentin expression and normal karyotype (2n = 58). The viability remained high (> 87%) and constant from the fourth to the tenth passage ( p  > 0.05). The passages did not change the cell morphology and metabolic and growth rates. Moreover, cryopreservation did not affect most evaluated parameters; post-thawed cells maintained their viability, growth, metabolism, and apoptosis levels. Nevertheless, cryopreservation increased mitochondrial membrane permeability and cell population doubling time compared to non-cryopreserved cells ( p