Isolation of Primary Fibroblast Culture from Wildlife: the Panthera onca Case to Preserve a South American Endangered Species

Abstract

Cell line establishment of somatic cells is a valuable resource to preserve genetic material of rare, difficult-to-find, endangered and giant species like Jaguar (Panthera onca), the largest South American felid. This unit focuses on the isolation and culture of fibroblasts from Jaguar skin and muscle biopsies, and ear cartilage dissection immediately after death to preserve one of the several endangered species in this biome. These culture techniques enabled us to contribute 570 samples from 45 autochthonous and endangered species, including Jaguar. The fibroblasts obtained are a part of the Genetic Bank of Buenos Aires Zoo with the 6700 samples, including tissues such as muscle, ovarian, testicular, blood, fibroblast cultures, sperm, hair, and fluids and cells from 450 individuals of 87 different species. © 2016 by John Wiley & Sons, Inc.

Quantitative and descriptive histological aspects of jaguar (Panthera onca Linnaeus, 1758) ear skin as a step towards formation of biobanks

“…The data of in vitro culture presented a very similar cell growth pattern for both jaguars, where cellular growth from the explants occurred in 9 and 10 days. These data were similar to that obtained by Mestre‐Citrinovitz et al (), who that reported that cell growth occurred between 10 and 14 days for the yellow jaguar. Furthermore, for other species of felines such as the Bengal tiger ( Panthera tigris tigris ) and Siberian tiger ( Panthera tigris altaica ), cell growth occurred after about 5–12 days of in vitro culture (Guan et al, ; Liu, Guo, Liu, Guan, & Ma, ).…”

“…We described the histological aspects of ear skin of the yellow and black jaguars, as this information is essential for a more targeted application of skin cryopreservation and subsequent cellular recovery. The formation of skin banks is of growing interest in conserving genetic samples of wild felids (Guan et al, ; León‐Quinto et al, ), especially jaguars (Mestre‐Citrinovitz et al, ), aimed at ensuring the storage of valuable genetic material (Praxedes, Borges, Santos, & Pereira, ).…”

“…Four skin fragments per animal were incubated in 35‐mm Petri dishes with DMEM supplemented with 10% FBS and 2% penicillin, streptomycin and amphotericin solution, at 38.5°C under controlled atmosphere with 5% CO 2 and 95% air, according to the method described by Mestre‐Citrinovitz et al (). For the primary culture, 35‐mm diameter Petri dishes were used with 1.0 ml of the medium, and for the subcultures, 60‐mm diameter Petri dishes were used with 4.0 ml of the medium.…”

“…Given the ecological and scientific importance of the species (Morato, Ferraz, Paula, & Campos, ), conservation tools such as skin cryopreservation become urgently needed to help protect genetic material which undergoes a reduction in population sizes. The first work published on the recovery and culture of somatic tissues for jaguars is the study conducted by Mestre‐Citrinovitz, Sestelo, Ceballos, Baranao, and Saragueta (). In this study, the authors emphasized only the description of the technical aspects for the recovery and culture of somatic tissues of jaguars, without presenting specifics such as cellular confluence and histology of the skin.…”

Isolation and preservation of multipotent mesenchymal stem cells from bone marrow of Arabian leopard (<i>Panthera pradus nimr</i>)

“…Cellular reprogramming for generating induced pluripotent stem cells (iPSCs) was recently adapted to reprogram diverse rare animal species, in an effort toward using this technology for preservation of the genetic material of endangered species. Successful trials resulting in generation of iPSCs from cryopreserved cell lines derived from populations of endangered Mandrillus leucophaeus, northern white rhinoceros, snow leopard were preformed (Leon-Quinto et al, 2009;Ben-Nun et al, 2011;Verma et al, 2012;Mestre-Citrinovitz et al, 2016). Somatic cells were reportedly used to clone a wild cat (Gómez et al, 2004).…”