An efficient method of generating skin fibroblast cells for cell banking

REKHA SHARMA; HIMANI SHARMA; SONIKA AHLAWAT; M S TANTIA

doi:10.56093/ijans.v88i8.82937

Authors

REKHA SHARMA Principal Scientist, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana 132 001 India
HIMANI SHARMA Senior Research Fellow, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana 132 001 India
SONIKA AHLAWAT Scientist, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana 132 001 India
M S TANTIA Principal Scientist, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana 132 001 India

https://doi.org/10.56093/ijans.v88i8.82937

Keywords:

Buffalo, Epithelial cells, Explant culture, Genebank, Skin fibroblasts

Abstract

An optimum cell culture medium is used to generate fibroblast cells for cell banking. Here, we describe a method for obtaining higher number of pure fibroblast cells in much shorter duration from skin tissue explants. Two pronged strategy was used, first two different culture media were selected, one for preferential generation of fibroblast cells in primary culture (Human fibroblast specific media- HifibroXL) and another for their faster multiplication in secondary culture (DMEM + Ham's F12). Secondly, tissue explants were cultured not only once but up to six times increasing the generation of primary cells per se. This method initially standardized with buffalo (Bubalus bubalis) skin tissue explants works efficiently with camel (Camelus bactrianus) and horse (Equs caballus). Modified cell culture method increases the efficiency of establishing fibroblast cell banks by reducing the cost both in terms of consumables and human effort. It is well suited for today's fast-paced conservation laboratories.

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References

Freshney R I. 2011. Primary culture, pp 163–186. Culture of Animals Cells: A Manual of Basic Technique and Specialized Applications. (Ed.) Freshney R I. Wiley-Liss Inc, New York. DOI: https://doi.org/10.1002/9780470649367.ch11

Hosseini S M, Moulavi F, Foruzanfar M, Hajian M, Abedi P, Rezazade-Valojerdi M, Parivar K, Shahverdi A H and Nasr- Esfahani M H. 2008. Effect of donor cell type and gender on the efficiency of in vitro sheep somatic cell cloning. Small Ruminant Research 78(1–3): 162–68. DOI: https://doi.org/10.1016/j.smallrumres.2008.06.004

Hui W, Xiangchen Li, Changli Li, Wenxiu Z, Weijun G and Yuehui Ma. 2012. Establishment and biological characteristics of Qing Kedan chicken embryonic fibroblast line for genetic resource conservation. Journal of Animal and Veterinary Advances 11: 4374–81.

Huschtscha L I, Napier C E, Noble J R, Bower K, Au A Y and Campbell H G. 2012. Enhance disolation of fibroblasts from human skin explants. Biotechniques 53: 239–44. DOI: https://doi.org/10.2144/0000113939

Liu C, Guo Y, Lu T, Li X, Guan W and Ma Y. 2014. Establishment and genetic characteristics analysis of in vitro culture a fibroblast cell line derived from Wuzhishan miniature pig. Cryobiology 68(2): 281–87. DOI: https://doi.org/10.1016/j.cryobiol.2014.02.009

Mehrabani D, Mahboobi R, Dianatpour M, Zare S, Tamadon A and Hosseini S E. 2014. Establishment, culture and characterization of Guinea pig fetal fibroblast cells. Veterinary Medicine International. Article ID #510328. DOI: https://doi.org/10.1155/2014/510328

Mehrabani D, Tajedini M, Tamadon A, Dianatpour M, Zare S and Rahmanifar F. 2016. Establishment, characterization and cryopreservation of Fars native goat fetal fibroblast cell lines. Asian Pacific Journal of Reproduction 5: 247–51. DOI: https://doi.org/10.1016/j.apjr.2016.04.013

Pei P, He X, Zhao Y, Zhu Z, Kan J, Guan W and Ma Y. 2015. Establishment and characterization of a fibroblast cell line derived from Gansu alpine fine-wool sheep. Biocell 39(1): 8– 13.

Singh M and Sharma A. 2011. Outgrowth of fibroblast cells from goat skin explants in three different culture media and the establishment of cell lines. In Vitro Cellular and Developmental Biology – Animal 47(2): 83–88. DOI: https://doi.org/10.1007/s11626-010-9373-4

Singh M, Sharma A K and Yadav P. 2011. Characterization of gsf289: a fibroblast cell line derived from goat ear skin explants. Journal of Biotech Research 3(1): 1–6.

Tracy L E, Minasian R A and Caterson E J. 2016. Extracellular matrix and dermal fibroblast function in the healing wound. Advances in Wound Care 5(3): 119–36. DOI: https://doi.org/10.1089/wound.2014.0561

Yi F, Liu G H and Izpisua Belmonte J C. 2012. Rejuvenating liver and pancreas through cell transdifferentiation. Cell Research 22: 616–19. DOI: https://doi.org/10.1038/cr.2012.33