In vitro culture of somatic cells derived from ear tissue of collared peccary (Pecari tajacu Linnaeus, 1758) in medium with different requirements

Abstract

The maintenance of metabolic activities during the in vitro culture of somatic cells of wild animals, especially collared peccary (Pecari tajacu), is an interesting step in conservation of these cells for the use in nuclear transfer. In this context, it is necessary to optimize the culture conditions of somatic cells by the establishment of appropriate supplementation to the media. Therefore, this study aimed to analyze the composition of the culture means of somatic cell derived from ear tissue of collared peccaries, evaluating concentrations of fetal bovine serum (FBS; 10% vs. 20%) and epidermal growth factor (EGF; 5ng/mL vs. 10ng/mL). Tissues were submitted to primary culture and subcultures for 40 days and cells were analyzed for morphology, adhesion, subconfluence, and proliferative activity to develop the growth curve and to determine the population doubling time (PDT), viability, and functional/metabolic activity. No difference was observed between the concentrations of FBS for several parameters, except for viability [FBS10: 85.6% vs. FBS20: 98.2%], PDT [FBS10: 155.4h vs. 77.2h], and functional/metabolic assay [FBS10: 0.57-0.55 vs. FBS20: 0.82-0.99 (D5-D7)]. For the EGF in culture, no difference was observed in the evaluated parameters. In all experiments, the growth curves were typical S-shape and the cells passed through a lag, logarithmic, and plateau phase. In conclusion, 20% FBS is suitable for the recovery of somatic cells; nevertheless, EGF does not improve the quality of growing these cells. To our knowledge, this is the first study culturing somatic cells of collared peccaries.

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

“…In this sense, studies related to the conservation of the collared peccary have been intensified, especially aimed at improving the techniques related to the preservation of somatic samples. Using this study, we established a culture condition for explants derived from the skin of adult collared peccaries (Santos et al, 2016) and developed a protocol for cryopreservation (Borges et al, 2017(Borges et al, , 2018a(Borges et al, , 2018b) and refrigeration of these explants (Queiroz Neta et al, 2018). In order to conduct the cloning experiments on this species by a somatic cell nuclear transfer, as well as to produce induced pluripotent cells, it is necessary to establish properly characterized cell lines.…”

Peer Review #4 of "Isolation, characterization, and cryopreservation of collared peccary skin-derived fibroblast cell lines (v0.1)"

“…Manuscript to be reviewed environment of the cells (Guo et al, 2018), we observed previously (Santos et al, 2016) that the medium for growth of somatic cells derived from collared peccaries was DMEM with 10% FBS and 2% antibiotic-antimycotic solution.…”

Combination of ethylene glycol with sucrose increases survival rate after vitrification of somatic tissue of collared peccaries (Pecari tajacu Linnaeus, 1758)

“…Initially, in the management of systems of the collared peccaries, their identification is recorded by ear sections and these fragments (1-2cm 2 ) were recovered for the experiment. Then, skin tissues were transported to the laboratory in Dulbecco modified Eagle medium (DMEM) with 2.2g/L sodium bicarbonate and 2% antibiotic-antimycotic solution at 37°C for 30 min, according to Santos et al (2016). At the laboratory, the tissue fragments were washed in 70% ethanol and DMEM.…”