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

Overview

Journal Cells

Publisher MDPI

Specialties Biochemistry
Biophysics
Cell Biology
Molecular Biology

Date 2021 Nov 27

PMID 34831260

Citations 3

Authors

Dalanda Wanes

Hassan Y Naim

Franziska Dengler

Affiliations

Soon will be listed here.

Abstract

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 . 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.

Citing Articles

Secretome of Adipose-Derived Stem Cells Cultured in Platelet Lysate Improves Migration and Viability of Keratinocytes.

Hermann M, Peddi A, Gerhards A, Schmid R, Schmitz D, Arkudas A Int J Mol Sci. 2023; 24(4).

PMID: 36834932 PMC: 9962933. DOI: 10.3390/ijms24043522.

The dark side of foetal bovine serum in extracellular vesicle studies.

Urzi O, Olofsson Bagge R, Crescitelli R J Extracell Vesicles. 2022; 11(10):e12271.

PMID: 36214482 PMC: 9549727. DOI: 10.1002/jev2.12271.

A Never-Ending Journey in Search for Novel Cell Biology Techniques.

Kalyuzhny A Cells. 2022; 11(9).

PMID: 35563699 PMC: 9101893. DOI: 10.3390/cells11091393.

References

Rauch C, Feifel E, Amann E, Spotl H, Schennach H, Pfaller W . Alternatives to the use of fetal bovine serum: human platelet lysates as a serum substitute in cell culture media. ALTEX. 2011; 28(4):305-16. DOI: 10.14573/altex.2011.4.305. View

Mirabet V, Solves P, Minana M, Encabo A, Carbonell-Uberos F, Blanquer A . Human platelet lysate enhances the proliferative activity of cultured human fibroblast-like cells from different tissues. Cell Tissue Bank. 2007; 9(1):1-10. DOI: 10.1007/s10561-007-9048-x. View

Nepal N, Arthur S, Sundaram U . Unique Regulation of Na-K-ATPase during Growth and Maturation of Intestinal Epithelial Cells. Cells. 2019; 8(6). PMC: 6628168. DOI: 10.3390/cells8060593. View

Shah P, Jogani V, Bagchi T, Misra A . Role of Caco-2 cell monolayers in prediction of intestinal drug absorption. Biotechnol Prog. 2006; 22(1):186-98. DOI: 10.1021/bp050208u. View

Henschler R, Gabriel C, Schallmoser K, Burnouf T, Koh M . Human platelet lysate current standards and future developments. Transfusion. 2019; 59(4):1407-1413. DOI: 10.1111/trf.15174. View

Hagen A, Lehmann H, Aurich S, Bauer N, Melzer M, Moellerberndt J . Scalable Production of Equine Platelet Lysate for Multipotent Mesenchymal Stromal Cell Culture. Front Bioeng Biotechnol. 2021; 8:613621. PMC: 7859354. DOI: 10.3389/fbioe.2020.613621. View

Hashimoto K, Shimizu M . Epithelial properties of human intestinal Caco-2 cells cultured in a serum-free medium. Cytotechnology. 1993; 13(3):175-84. DOI: 10.1007/BF00749813. View

Fazzina R, Iudicone P, Mariotti A, Fioravanti D, Procoli A, Cicchetti E . Culture of human cell lines by a pathogen-inactivated human platelet lysate. Cytotechnology. 2015; 68(4):1185-95. PMC: 4960166. DOI: 10.1007/s10616-015-9878-5. View

Alfalah M, Jacob R, Preuss U, Zimmer K, Naim H, Naim H . O-linked glycans mediate apical sorting of human intestinal sucrase-isomaltase through association with lipid rafts. Curr Biol. 1999; 9(11):593-6. DOI: 10.1016/s0960-9822(99)80263-2. View

10.

van der Valk J, Bieback K, Buta C, Cochrane B, Dirks W, Fu J . Fetal Bovine Serum (FBS): Past - Present - Future. ALTEX. 2017; 35(1):99-118. DOI: 10.14573/altex.1705101. View

11.

Thieme D, Reuland L, Lindl T, Kruse F, Fuchsluger T . Optimized human platelet lysate as novel basis for a serum-, xeno-, and additive-free corneal endothelial cell and tissue culture. J Tissue Eng Regen Med. 2017; 12(2):557-564. DOI: 10.1002/term.2574. View

12.

Naim H, Sterchi E, Lentze M . Biosynthesis of the human sucrase-isomaltase complex. Differential O-glycosylation of the sucrase subunit correlates with its position within the enzyme complex. J Biol Chem. 1988; 263(15):7242-53. View

13.

Gstraunthaler G . Alternatives to the use of fetal bovine serum: serum-free cell culture. ALTEX. 2003; 20(4):275-81. View

14.

Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A . Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002; 3(7):RESEARCH0034. PMC: 126239. DOI: 10.1186/gb-2002-3-7-research0034. View

15.

Burnouf T, Strunk D, Koh M, Schallmoser K . Human platelet lysate: Replacing fetal bovine serum as a gold standard for human cell propagation?. Biomaterials. 2015; 76:371-87. DOI: 10.1016/j.biomaterials.2015.10.065. View

16.

Shimada Y, Inomata M, Suzuki H, Hayashi M, Waheed A, Ohno-Iwashita Y . Separation of a cholesterol-enriched microdomain involved in T-cell signal transduction. FEBS J. 2005; 272(21):5454-63. DOI: 10.1111/j.1742-4658.2005.04938.x. View

17.

Schmitz J, PREISER H, Maestracci D, Ghosh B, Cerda J, CRANE R . Purification of the human intestinal brush border membrane. Biochim Biophys Acta. 1973; 323(1):98-112. DOI: 10.1016/0005-2736(73)90434-3. View

18.

ZWEIBAUM A, Triadou N, Kedinger M, Augeron C, Pinto M, Rousset M . Sucrase-isomaltase: a marker of foetal and malignant epithelial cells of the human colon. Int J Cancer. 1983; 32(4):407-12. DOI: 10.1002/ijc.2910320403. View

19.

Johansson L, Klinth J, Holmqvist O, Ohlson S . Platelet lysate: a replacement for fetal bovine serum in animal cell culture?. Cytotechnology. 2008; 42(2):67-74. PMC: 3449796. DOI: 10.1023/B:CYTO.0000009820.72920.cf. View

20.

Ferruzza S, Rossi C, Sambuy Y, Scarino M . Serum-reduced and serum-free media for differentiation of Caco-2 cells. ALTEX. 2013; 30(2):159-68. DOI: 10.14573/altex.2013.2.159. View