Isolation, characterization, and ecotoxicological application of marine mammal skin fibroblast cultures

Establishment and Characterization of a Primary Fibroblast Cell Culture from the Amazonian Manatee (Trichechus inunguis)

“…Cell cultures have diverse applications and are extremely important for the conservation of threatened species [ 33 , 34 , 35 ], such as manatee species. The establishment and characterization of primary cell lines are an important initial step for research in biotechnological and ecotoxicological applications to address crucial questions about conserving the genetic heritage of wildlife, the susceptibility of these species in the environment, and how they are affected [ 17 ]. From a biotechnological perspective, establishing and characterizing manatee cell lines can make them an effective resource for somatic cell nuclear transfer experiments [ 12 , 36 ], enhancing their efficiency in responding to cell viability, both for their own maintenance through passages, growth phases, cryopreservation, and recovery after thawing, as well as for the future use of these induced cell lines in toxicological analyses and gene therapies [ 13 , 19 , 37 ].…”

“…From an ecotoxicological perspective, it has become important to observe how TINsf can respond to environmental conditions in aquatic environments. This has already been addressed in various aquatic mammals, mainly to understand how the effects of aquatic pollution directly impact these species [ 17 , 18 , 21 ], such as studies conducted on cells obtained from fin whales ( Balaenoptera physalus ) [ 40 ], humpback whales ( Megaptera novaeangliae ) [ 18 ], and killer whales ( Orcinus orca ) [ 16 ], which have shown cytotoxic and genotoxic effects of environmental pollutants on the cell lines of these species.…”

“…The acquisition of somatic cells as fibroblast lines has been widely established in different representatives of global wildlife, mostly consisting of vertebrates such as mammals, birds, amphibians, and reptiles, especially those facing the threat of extinction [ 14 , 15 ]. Globally, these studies have been increasing in aquatic mammals in recent years [ 16 , 17 , 18 , 19 , 20 , 21 ], validating the importance of cryopreservation studies, especially for free-living animals susceptible to environmental changes. Mammalian cell culture can be applied in a variety of cellular assays to investigate morphology, protein expression, cell growth, differentiation, apoptosis, and toxicity in different environments, provided that the quality of cell line characterization is ensured [ 13 ].…”

Highly efficient isolation and 3D printing of fibroblasts for cultured meat production

“…The enzymatic digestion method first degrades the ECM in the skin tissue using proteases such as collagenase, thus usually achieving relatively high isolation efficiency and cell number of fibroblast (Wang et al, 2004). However, compared to humans and mice, the skin of livestock such as pigs and cows is thicker and more compact, so conventional enzymatic digestion processes may not be effective in isolating fibroblasts from livestock (Boroda et al, 2020). In addition to tissue explanting and enzymatic digestion, physical friction is another attractive method to promote cell release during the tissue dissociation process.…”

The initial steps toward the formation of somatic tissue banks and cell cultures derived from captive Antillean manatee (Trichechus manatus manatus) skin biopsies

“…Given this scenario, one strategy for the conservation of marine species is to establish in vitro somatic cell culture systems. These instruments are important tools for studies in the fields of cell biology, physiology, ecology, molecular biology, genetics, and ecotoxicological applications in these species (Boroda et al, 2020). Moreover, as in other marine and wild species, these cells can be valuable tools in the study of cellular reprogramming (Robles et al, 2017; Verma et al, 2012), aiming at the production of induced pluripotent cells for the generation of gametes.…”