Establishment, characterization, and cryopreservation of cell lines derived from red-rumped agouti (Dasyprocta leporina Linnaeus, 1758) – A study in a wild rodent

Praxedes, Érika Almeida; Silva, Maria Bárbara; Oliveira, Lhara Ricarliany Medeiros de; Viana, João Vitor da Silva; Silva, Alexandre Rodrigues; Oliveira, Moacir Franco de; Pereira, Alexsandra Fernandes

doi:10.1016/j.cryobiol.2020.12.006

Cited by 14 publications

(7 citation statements)

References 34 publications

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“…Thus, the explant isolation method is the most utilized in wild mammals due to its lower cost and lesser capacity to damage the fragment. The isolation by explants showed success in several wild felids, such as jaguar (Panthera onca) [12], and wild rodents, such as red-rumped agouti (Dasyprocta leporina) [13], and large Japanese field mice (Apodemus speciosus) [14] with cells recovered after 4 or 11 days.…”

Section: Isolation Techniques and Identification Of Fibroblastsmentioning

confidence: 99%

“…However, molecular analysis has been shown to be more reliable for this purpose [15]. Some methods have been applied, such as the florescence in Theriogenology -Recent Advances in the Field immunocytochemistry by an antibody specific for cell surface protein conjugate with fluorophore [13]. The quantitative real-time polymerase chain reaction (qRT-PCR) by gene expression has shown great results in several wild mammals [16].…”

Section: Isolation Techniques and Identification Of Fibroblastsmentioning

confidence: 99%

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Strategies for the Establishment of Fibroblastic Lines for the Conservation of Wild Mammals

Fernandes Pereira,

Ricarliany Medeiros de Oliveira,

Vitorino Costa de Aquino

et al. 2023

Veterinary Medicine and Science

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The loss of wild biodiversity has encouraged the development of fibroblastic lines, mainly fibroblasts derived from skin, which can be interesting tools for the conservation of wild mammals. These biological samples, when properly well-established, are essential elements for the reproduction of species through their use in cloning by somatic cell nuclear transfer and induction of cells to pluripotency. In general, the establishment of fibroblastic lines involves the following strategies: (i) cell isolation techniques and identification of fibroblasts; (ii) conditions for in vitro culture of fibroblasts; (iii) conditions for cryopreservation of fibroblasts; and (iv) nuclear reprogramming studies. At each stage, species-specific factors are involved, and determining these lines in the species of interest represents the first step toward its successful use for animal conservation. Therefore, this chapter discusses the stages and parameters involved in the strategies for establishing fibroblastic lines, delving into the main technical aspects and results obtained from the use of these cells in recent years in wild mammals.

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Section: Isolation Techniques and Identification Of Fibroblastsmentioning

confidence: 99%

Section: Isolation Techniques and Identification Of Fibroblastsmentioning

confidence: 99%

Strategies for the Establishment of Fibroblastic Lines for the Conservation of Wild Mammals

Fernandes Pereira,

Ricarliany Medeiros de Oliveira,

Vitorino Costa de Aquino

et al. 2023

Veterinary Medicine and Science

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“…In addition, Naaldijk et al reported that the cryopreservation of human fibroblasts in an adherent state significantly diminishes their subsequent growth potential owing to the fact that adherent cells are more vulnerable to the mechanical stress induced by cryopreservation [ 4 ]. Praxedes et al indicated that compared to non-cryopreserved rodent fibroblasts, the cryopreserved fibroblasts exhibited lower mitochondrial membrane potential and viability [ 9 ].…”

Section: Introductionmentioning

confidence: 99%

Evaluating the Effects of Cryopreservation on the Viability and Gene Expression of Porcine-Ear-Skin Fibroblasts

Cao

Xie

Wang

et al. 2023

Genes

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Owing to the inherent heterogeneity and plasticity of fibroblasts, they are considered as the conventional biological resources for basic and clinical medical research. Thus, it is essential to generate knowledge about the establishment of fibroblast cultures and the effects of cryopreservation processes on their biological characteristics. Since the pig (Sus scrofa) possesses numerous genetic, physiological, and anatomical similarities with humans, porcine fibroblasts are naturally regarded as useful analogues of human fibroblasts. Nonetheless, less attention has been given to the alterations in viability and gene expression of cryopreserved porcine fibroblasts. In this study, we aimed to obtain fibroblasts from porcine ear skin and evaluate the effects of cryopreservation on the cell survival, proliferation, and gene expression profiles of the fibroblasts by trypan-blue-staining assay, Cell Counting kit-8 (CCK-8) assay, and RNA-sequencing analysis, respectively. Our results suggested that morphologically stable fibroblast cultures can be constructed from pig-ear skin. The post-thaw survival rate of the cryopreserved fibroblasts at 0 h and 24 h was over 90%. The proliferative activity of the cryopreserved fibroblasts was similar to that of the non-cryopreserved fibroblasts after 7 days of in vitro culture, which suggested that cryopreservation did not influence the viability. The RNA-sequencing analysis indicated that this should be attributed to the 867 differentially expressed genes (DGEs) identified, which are involved in molecular process related to cell recovery and survival after cryo-stimulation. In addition, eight important DEGs BMP2, GDF15, EREG, AREG, HBEGF, LIF, IL-6, and HOX-7 could potentially be applied to improve the efficiency of fibroblast cryopreservation, but comprehensive and systematic studies on understanding the underlying mechanisms responsible for their modulatory roles are urgently needed.

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“…Although classified as of “Least Concern” regarding the threat of extinction (Emmons & Reid, 2016), hunting and habitat fragmentation have resulted in a population decrease of the species in some regions (Ferreguetti et al, 2018). In this scenario, different conservation strategies involving assisted reproductive techniques have been proposed for males (Castelo et al, 2015), females (Praxedes et al, 2018a), as well as the use of somatic tissues (Costa et al, 2020), and cells (Praxedes et al, 2021) of red‐rumped agouti.…”

Section: Introductionmentioning

confidence: 99%

“…been proposed for males (Castelo et al, 2015), females (Praxedes et al, 2018a), as well as the use of somatic tissues (Costa et al, 2020), and cells (Praxedes et al, 2021) of red-rumped agouti.…”

mentioning

confidence: 99%

Synergistic effects of follicle‐stimulating hormone and epidermal growth factor on in vitro maturation and parthenogenetic development of red‐rumped agouti oocytes

Praxedes,

Santos,

de Oliveira

et al. 2023

Reprod Domestic Animals

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Although oocyte in vitro maturation (IVM) is routinely used for in vitro embryo production in mice and rats, its use in wild rodents remains unexplored. Evidence suggests that hormone and growth factor supplementation influence oocyte meiotic resumption. This study evaluated the synergistic effects of follicle‐stimulating hormone (FSH) and epidermal growth factor (EGF) on the IVM and parthenogenetic development of red‐rumped agouti oocytes. Initially, we evaluated the IVM rates, mature oocyte quality, oocyte morphometry, and early embryonic development during IVM in the presence of 10, 50, and 75 mIU/mL FSH. No differences among the FSH concentrations were observed for IVM rates, oocyte morphometry, cumulus cell expansion, and viability. Although oocytes matured with 50 mIU/mL FSH showed a higher rate of cumulus expansion index (CEI), only oocytes matured with 10 mIU/mL FSH resulted in morulae after chemical activation (7.9% ± 4.2%). Thus, 10 mIU/mL FSH was used for further experiments. We subsequently evaluated the synergistic effects of 10, 50, and 100 ng/mL EGF and 10 mIU/mL FSH on the same parameters. No differences among the groups were observed in IVM rates, oocyte morphometry, and cumulus viability. Nevertheless, FSH with 10 ng/mL EGF showed a CEI superior to that of the other groups. Furthermore, oocytes matured with FSH alone or with both FSH and 10 or 50 ng/mL EGF developed morulae after activation (5.8%–8.3%). In conclusion, oocytes matured with 10 mIU/mL FSH and 10 ng/mL EGF are recommended for use in red‐rumped agouti oocyte IVM, as they positively influence embryonic development.

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Establishment, characterization, and cryopreservation of cell lines derived from red-rumped agouti (Dasyprocta leporina Linnaeus, 1758) – A study in a wild rodent

Cited by 14 publications

References 34 publications

Strategies for the Establishment of Fibroblastic Lines for the Conservation of Wild Mammals

Strategies for the Establishment of Fibroblastic Lines for the Conservation of Wild Mammals

Evaluating the Effects of Cryopreservation on the Viability and Gene Expression of Porcine-Ear-Skin Fibroblasts

Synergistic effects of follicle‐stimulating hormone and epidermal growth factor on in vitro maturation and parthenogenetic development of red‐rumped agouti oocytes

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