Isolation, characterization, and cryopreservation of collared peccary skin-derived fibroblast cell lines.

摘要: Background Biobanking of cell lines is a promising tool support for wildlife conservation. In particular, the ability to preserve fibroblast derived from collared peccaries significance as these wild mammals are unique Americas and play large role in maintaining ecosystem. We identified peccary fibroblasts by immunofluorescence evaluated their morphology, growth adherence capacity. Further, we monitored viability metabolic activity determine effects passage number cryopreservation on establishment lines. Methods Skin biopsies were collected peripheral ear region five adult animals captivity. Initially, cells isolated fragments cultured Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum 2% antibiotic-antimycotic solution under controlled atmosphere (38.5 °C, 5% CO2). maintenance primary capacity explants, explants subconfluence, absence contamination. Moreover, immunofluorescence. Additionally, evaluate influence passages (first, third tenth passage) lines, analysed viability, activity, population doubling time (PDT), levels reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm). Results All (20/20) adhered dish 2.4 days ± 0.5 around 4.6 0.7, subconfluence was observed within 7.8 1.0. morphology immunocytochemistry analyses, which presented oval nuclei, fusiform shape positive vimentin staining. No contamination after culture without antibiotics antifungals 30 days. While there no difference (first vs. third: P = 0.98; first tenth: 0.76; 0.85), found be reduced (23.2 12.1%) when compared that (100.0 24.4%, 0.006). did not (P 0.11), 0.77), or PDT 0.11). Nevertheless, greater ΔΨm 0.0001) cryopreserved (2.12 0.14) non-cryopreserved (1.00 0.05). showed intracellular ROS thawing (1.69 0.38 1.00 0.22, 0.04). Conclusions This study report isolation, characterization peccaries. adherent cultures efficient obtaining fibroblasts, can used donor nuclei cloning other applications.