Simple Method for Establishing Primary Leporidae Skin Fibroblast Cultures

Santos, Fábio A. Abade dos; Carvalho, Carina L.; Almeida, Isabel Tavares de; Fagulha, Teresa; Rammos, Fernanda; Barros, Sílvia C.; Henriques, M. J.; Luís, Tiago; Duarte, Margarida

doi:10.3390/cells10082100

Cited by 10 publications

(3 citation statements)

References 18 publications

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“…Specifically, regarding the methods of cell isolation, the three cited methods can be conducted alone or combined. The mechanical dissolution in combination with dissolution enzymatic was conducted in Cheetah (Acinonyx jubatus) [9], Asian elephant (Elephas maximus) [10], Iberian hare (Lepus granatensis), and European rabbit (Oryctolagus cuniculus) [11] with cells recovered after 4-7 days. However, enzymatic, and mechanical methods can result in damage, reducing the cell viability if inconsistently utilized.…”

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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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%

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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“…Historically, epithelia have proved more di cult to culture than many other cell types, and only very limited culture was possible [12]. However, primary cells retain the same characteristics as those of the original tissue, thus facilitating studies in many research areas, including medical and cell biology research, that otherwise would be far more complicated [13].…”

Section: Introductionmentioning

confidence: 99%

A method for isolating and culturing ectopic epithelial and stromal cells to study human adenomyosis

Fang,

Wang,

et al. 2023

Arch Gynecol Obstet

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Although adenomyosis is a common and benign gynecological disease, the speci c pathogenesis of this condition has yet to be fully elucidated. It is di cult to culture primary cells of the ectopic endometrial epithelia and stroma from human adenomyosis lesions. Most previous of studies on adenomyosis were based on primary eutopic endometrium cells. However, as yet, no e cient protocols have been developed for the isolation, culture or puri cation of primary ectopic epithelial and stromal cells from human adenomyosis lesions. Therefore, the present study aimed to develop an e cient protocol for the isolation and culture of primary ectopic epithelial and stromal cells from human adenomyosis lesions. MethodsIn the present study, we aimed to obtain ectopic endometrium tissue from human adenomyosis foci and use a simple and operable type I collagenase digestion method for primary culture. Cells were isolated by sterile cell strainer ltration and ow cytometry was performed to identify, purify and evaluate the viability of isolated ectopic endometrial cells. ResultsUsing our method, we successfully isolated and cultured highly puri ed and active ectopic endometrial epithelial and stromal cells from human adenomyosis foci. Ep-CAM was expressed in ectopic epithelial cells of human adenomyosis with a purity of 93.74% and a viability of 80.58%. In addition, CD10 were robustly expressed by ectopic stromal cells in human adenomyosis. Cellular purity and viability were determined to be 96.37% and 93.49%, respectively. ConclusionOur method provides a new experimental model for studying the molecular pathogenesis of human adenomyosis.

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“…This is a very important finding that may help eliminate the use of the controversial FCS as a cell culture additive. It will be interesting to see if using PL can be also used to improve cell culture of primary fibroblasts from leporid species: Abade dos Santos et al described a simple, inexpensive technique [8] to generate the primary cell culture of fibroblasts from leporid species (rabbits and hares), which are widely used for biomedical research. Their novel technique allows the achievement of high viability and homogeneity of fibroblasts.…”

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confidence: 99%