Determining Influence of Culture Media and Dose-Dependent Supplementation with Basic Fibroblast Growth Factor on the Ex Vivo Proliferative Activity of Domestic Cat Dermal Fibroblasts in Terms of Their Suitability for Cell Banking and Somatic Cell Cloning of Felids

Wiesława Młodawska , Patrycja Mrowiec , Beata Grabowska , Joanna Waliszewska , Joanna Kochan , Agnieszka Nowak , Anna Migdał , Wojciech Niżański , Sylwia Prochowska , Agnieszka Partyka , Marcin Pałys , Teresa Grega , Józef Skotnicki

Abstract

Dermal fibroblasts are commonly used as donors of genetic material for somatic cell nuclear transfer in mammals. Basic fibroblast growth factor (bFGF) is a cytokine that regulates proliferation and differentiation of different cell types. The study was aimed at optimizing the cell culture protocol for cat dermal fibroblasts by assessing the influence of culture media and different doses of bFGF on proliferation of fibroblasts and their viability in terms of cell banking and somatic cloning of felids. In Experiment I, skin biopsies of domestic cats were cultured in DMEM (D) and/or DMEM/F12 (F), both supplemented with 5 ng bFGF/ml (D-5, F-5, respectively). After the primary culture reached ~80% of confluency, the cells were passaged (3–4 times) and cultured in media with (D-5, F-5) or without (D-0, F-0) bFGF. To determine the optimal doses of bFGF, in Experiment II, secondary fibroblasts were cultured in DMEM with 0 (D-0), 2.5 (D-2.5), 5 (D-5) or 10 (D-10) ng bFGF/ml. The results showed that in D-5 the cells proliferated faster than in D-0, F-5 and F-0. Due to their poor proliferation, passages IV were not performed for cells cultured in F-0. In experiment II, a dose-dependent effect of bFGF on proliferation of cat dermal fibroblasts was found. In D-5 and D-10, the cells exhibited higher (P<0.05) proliferation compared with D-0. In D-2.5 the cells showed a tendency to proliferate slower than in D-5 and D-10 and at the same faster than in D-0. In conclusion. DMEM supplemented with bFGF provides better proliferation of domestic cat dermal fibroblasts culture than DMEM/F12. Supplementation of culture medium with bFGF has a beneficial effect on cat dermal fibroblast proliferation and could be recommended for addition to culture media.
Author Wiesława Młodawska (FoAS / IoVS)
Wiesława Młodawska,,
- Institute of Veterinary Sciences
, Patrycja Mrowiec (FoAS / IoAS)
Patrycja Mrowiec,,
- Institute of Animal Science
, Beata Grabowska (FoAS / IoVS)
Beata Grabowska,,
- Institute of Veterinary Sciences
, Joanna Waliszewska (FoAS / IoVS)
Joanna Waliszewska,,
- Institute of Veterinary Sciences
, Joanna Kochan (FoAS / IoVS)
Joanna Kochan,,
- Institute of Veterinary Sciences
, Agnieszka Nowak (FoAS / IoVS)
Agnieszka Nowak,,
- Institute of Veterinary Sciences
, Anna Migdał (FoAS / IoVS)
Anna Migdał,,
- Institute of Veterinary Sciences
, Wojciech Niżański
Wojciech Niżański,,
-
, Sylwia Prochowska
Sylwia Prochowska,,
-
, Agnieszka Partyka
Agnieszka Partyka,,
-
et al.`
Journal seriesAnnals of Animal Science, ISSN 1642-3402, e-ISSN 2300-8733, (N/A 100 pkt)
Issue year2019
Vol19
No2
Pages359-372
Publication size in sheets0.65
Keywords in Englishcat; skin; fibroblasts; cell culture; basic fibroblast growth factor
ASJC Classification3403 Food Animals; 3404 Small Animals; 1103 Animal Science and Zoology
DOIDOI:10.2478/aoas-2018-0057
URL https://content.sciendo.com/view/journals/aoas/19/2/article-p359.xml
Internal identifierWHiBZ/2019/29
Languageen angielski
Score (nominal)100
Score sourcejournalList
Publication indicators WoS Citations = 0; Scopus SNIP (Source Normalised Impact per Paper): 2018 = 0.869; WoS Impact Factor: 2018 = 1.515 (2) - 2018=1.246 (5)
Citation count*
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FinansowanieNCBiR PBS3/B8/16/2015
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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