Proliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum

Cell lines are widely used as in vitro model systems and substitute for animal experiments. The frequently used Caco-2 cell line is considered to reflect characteristics of differentiated intestinal epithelium. However, the need to culture the cells with fetal calf serum (FCS) induces a high variabi...

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Autores principales:	Dalanda Wanes, Hassan Y. Naim, Franziska Dengler
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	3R cell culture differentiation FCS hPL gene expression Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/eed297bc71374daeb1d35bfe62330be5
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spelling	oai:doaj.org-article:eed297bc71374daeb1d35bfe62330be52021-11-25T17:10:39ZProliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum10.3390/cells101130382073-4409https://doaj.org/article/eed297bc71374daeb1d35bfe62330be52021-11-01T00:00:00Zhttps://www.mdpi.com/2073-4409/10/11/3038https://doaj.org/toc/2073-4409Cell lines are widely used as in vitro model systems and substitute for animal experiments. The frequently used Caco-2 cell line is considered to reflect characteristics of differentiated intestinal epithelium. However, the need to culture the cells with fetal calf serum (FCS) induces a high variability, risk of contamination and is ethically disputed. We tested the culture of Caco-2 cells with human platelet lysate (PL) instead of FCS. We compared cell viability and differentiation by measuring ATP levels, gene and protein expression of specific markers in total cell extracts, brush border membrane vesicles (BBM) and lipid rafts (LR). Cell viability was slightly enhanced in cells grown with PL compared to FCS. The cells differentiated to an intestinal phenotype like the cells cultured in FCS, as indicated by the similar gene expression levels of hexose and protein transport proteins and the structural protein <i>VILLIN</i>. BBM showed a comparable distribution of the intestinal hydrolases, indicating a maintained cell membrane polarity. The distribution of the marker protein FLOTILLIN-2 in LR was also similar. We conclude that PL is an exquisite and suitable replacement for FCS in the culture of Caco-2 cells that can eliminate many disadvantages incurred due to the use of FCS.Dalanda WanesHassan Y. NaimFranziska DenglerMDPI AGarticle3Rcell culturedifferentiationFCShPLgene expressionBiology (General)QH301-705.5ENCells, Vol 10, Iss 3038, p 3038 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	3R cell culture differentiation FCS hPL gene expression Biology (General) QH301-705.5
spellingShingle	3R cell culture differentiation FCS hPL gene expression Biology (General) QH301-705.5 Dalanda Wanes Hassan Y. Naim Franziska Dengler Proliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum
description	Cell lines are widely used as in vitro model systems and substitute for animal experiments. The frequently used Caco-2 cell line is considered to reflect characteristics of differentiated intestinal epithelium. However, the need to culture the cells with fetal calf serum (FCS) induces a high variability, risk of contamination and is ethically disputed. We tested the culture of Caco-2 cells with human platelet lysate (PL) instead of FCS. We compared cell viability and differentiation by measuring ATP levels, gene and protein expression of specific markers in total cell extracts, brush border membrane vesicles (BBM) and lipid rafts (LR). Cell viability was slightly enhanced in cells grown with PL compared to FCS. The cells differentiated to an intestinal phenotype like the cells cultured in FCS, as indicated by the similar gene expression levels of hexose and protein transport proteins and the structural protein <i>VILLIN</i>. BBM showed a comparable distribution of the intestinal hydrolases, indicating a maintained cell membrane polarity. The distribution of the marker protein FLOTILLIN-2 in LR was also similar. We conclude that PL is an exquisite and suitable replacement for FCS in the culture of Caco-2 cells that can eliminate many disadvantages incurred due to the use of FCS.
format	article
author	Dalanda Wanes Hassan Y. Naim Franziska Dengler
author_facet	Dalanda Wanes Hassan Y. Naim Franziska Dengler
author_sort	Dalanda Wanes
title	Proliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum
title_short	Proliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum
title_full	Proliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum
title_fullStr	Proliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum
title_full_unstemmed	Proliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum
title_sort	proliferation and differentiation of intestinal caco-2 cells are maintained in culture with human platelet lysate instead of fetal calf serum
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/eed297bc71374daeb1d35bfe62330be5
work_keys_str_mv	AT dalandawanes proliferationanddifferentiationofintestinalcaco2cellsaremaintainedinculturewithhumanplateletlysateinsteadoffetalcalfserum AT hassanynaim proliferationanddifferentiationofintestinalcaco2cellsaremaintainedinculturewithhumanplateletlysateinsteadoffetalcalfserum AT franziskadengler proliferationanddifferentiationofintestinalcaco2cellsaremaintainedinculturewithhumanplateletlysateinsteadoffetalcalfserum
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Proliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum

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