A Novel Epithelial and Fibroblastic Cell Isolation and Purification Method Using Primary Culture of Bovine Tongue Tissue

Cell lines provide a useful in vitro model to study in different biotechnological fields. The purity of the cell lines plays a pivotal role in research and production activities. This study aims to present a new method to establish pure cell lines by using bovine tongue tissue. For this purpose, the...

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Autor principal:	Sukran YILMAZ
Formato:	article
Lenguaje:	EN
Publicado:	University of Kafkas 2021
Materias:	primary cell culture fibroblast cell epithelial cell pure culture Veterinary medicine SF600-1100
Acceso en línea:	https://doaj.org/article/6b8d5a53b39f46dd9e90c8d6705c41f1
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spelling	oai:doaj.org-article:6b8d5a53b39f46dd9e90c8d6705c41f12021-12-03T07:50:10ZA Novel Epithelial and Fibroblastic Cell Isolation and Purification Method Using Primary Culture of Bovine Tongue Tissue10.9775/kvfd.2021.259931300-60451309-2251https://doaj.org/article/6b8d5a53b39f46dd9e90c8d6705c41f12021-11-01T00:00:00Zhttp://vetdergikafkas.org/uploads/pdf/pdf_KVFD_2846.pdfhttps://doaj.org/toc/1300-6045https://doaj.org/toc/1309-2251Cell lines provide a useful in vitro model to study in different biotechnological fields. The purity of the cell lines plays a pivotal role in research and production activities. This study aims to present a new method to establish pure cell lines by using bovine tongue tissue. For this purpose, the bovine tongue was obtained from the local slaughterhouse. After establishing primary cell culture, the cells were treated with EDTA (0.02%) for 3-5 min at 37°C. Primarily detached fibroblasts were collected into 5 mL EMEM (with 20% FBS), centrifuged and transferred to a new culture flask with EMEM (10% FCS) medium. The remainder cells in the primary flask were incubated for 8 hours with DMEM (10% FCS). Thereafter, the same process was applied with Na2EDDA (0.01%), and the cells were washed twice with DMEM (with 20% FBS). Reincubation was carried out with decreased FBS concentration (10%) and EGF (10 ng/mL) at 37°C, 5% CO2 in humidified air conditions. The same processes were repeated after 48 h. Conformational studies of pure cultures were done with immunostaining technique with anti-cytokeratin and antivimentin monoclonal antibodies where, after purification, fibroblasts displayed vimentin-positive and epithelial cells cytokeratin-positive. In conclusion, this study successfully demonstrated an easy, effective, and efficient method to generate pure novel cell lines from primary tissue culture or mixed cell cultures.Sukran YILMAZUniversity of Kafkasarticleprimary cell culturefibroblast cellepithelial cellpure cultureVeterinary medicineSF600-1100ENKafkas Universitesi Veteriner Fakultesi Dergisi, Vol 27, Iss 6, Pp 675-680 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	primary cell culture fibroblast cell epithelial cell pure culture Veterinary medicine SF600-1100
spellingShingle	primary cell culture fibroblast cell epithelial cell pure culture Veterinary medicine SF600-1100 Sukran YILMAZ A Novel Epithelial and Fibroblastic Cell Isolation and Purification Method Using Primary Culture of Bovine Tongue Tissue
description	Cell lines provide a useful in vitro model to study in different biotechnological fields. The purity of the cell lines plays a pivotal role in research and production activities. This study aims to present a new method to establish pure cell lines by using bovine tongue tissue. For this purpose, the bovine tongue was obtained from the local slaughterhouse. After establishing primary cell culture, the cells were treated with EDTA (0.02%) for 3-5 min at 37°C. Primarily detached fibroblasts were collected into 5 mL EMEM (with 20% FBS), centrifuged and transferred to a new culture flask with EMEM (10% FCS) medium. The remainder cells in the primary flask were incubated for 8 hours with DMEM (10% FCS). Thereafter, the same process was applied with Na2EDDA (0.01%), and the cells were washed twice with DMEM (with 20% FBS). Reincubation was carried out with decreased FBS concentration (10%) and EGF (10 ng/mL) at 37°C, 5% CO2 in humidified air conditions. The same processes were repeated after 48 h. Conformational studies of pure cultures were done with immunostaining technique with anti-cytokeratin and antivimentin monoclonal antibodies where, after purification, fibroblasts displayed vimentin-positive and epithelial cells cytokeratin-positive. In conclusion, this study successfully demonstrated an easy, effective, and efficient method to generate pure novel cell lines from primary tissue culture or mixed cell cultures.
format	article
author	Sukran YILMAZ
author_facet	Sukran YILMAZ
author_sort	Sukran YILMAZ
title	A Novel Epithelial and Fibroblastic Cell Isolation and Purification Method Using Primary Culture of Bovine Tongue Tissue
title_short	A Novel Epithelial and Fibroblastic Cell Isolation and Purification Method Using Primary Culture of Bovine Tongue Tissue
title_full	A Novel Epithelial and Fibroblastic Cell Isolation and Purification Method Using Primary Culture of Bovine Tongue Tissue
title_fullStr	A Novel Epithelial and Fibroblastic Cell Isolation and Purification Method Using Primary Culture of Bovine Tongue Tissue
title_full_unstemmed	A Novel Epithelial and Fibroblastic Cell Isolation and Purification Method Using Primary Culture of Bovine Tongue Tissue
title_sort	novel epithelial and fibroblastic cell isolation and purification method using primary culture of bovine tongue tissue
publisher	University of Kafkas
publishDate	2021
url	https://doaj.org/article/6b8d5a53b39f46dd9e90c8d6705c41f1
work_keys_str_mv	AT sukranyilmaz anovelepithelialandfibroblasticcellisolationandpurificationmethodusingprimarycultureofbovinetonguetissue AT sukranyilmaz novelepithelialandfibroblasticcellisolationandpurificationmethodusingprimarycultureofbovinetonguetissue
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A Novel Epithelial and Fibroblastic Cell Isolation and Purification Method Using Primary Culture of Bovine Tongue Tissue

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