Validation of in vitro assays in three-dimensional human dermal constructs

The International Journal of Artificial Organs
Ayesha IdreesJochen Salber

Abstract

Three-dimensional cell culture systems are urgently needed for cytocompatibility testing of biomaterials. This work aimed at the development of three-dimensional in vitro dermal skin models and their optimization for cytocompatibility evaluation. Initially "murine in vitro dermal construct" based on L929 cells was generated, leading to the development of "human in vitro dermal construct" consisting of normal human dermal fibroblasts in rat tail tendon collagen type I. To assess the viability of the cells, different assays CellTiter-Blue®, RealTime-Glo™ MT, and CellTiter-Glo® (Promega) were evaluated to optimize the best-suited assay to the respective cell type and three-dimensional system. Z-stack imaging (Live/Dead and Phalloidin/DAPI-Promokine) was performed to visualize normal human dermal fibroblasts inside matrix revealing filopodia-like morphology and a uniform distribution of normal human dermal fibroblasts in matrix. CellTiter-Glo was found to be the optimal cell viability assay among those analyzed. CellTiter-Blue reagent affected the cell morphology of normal human dermal fibroblasts (unlike L929), suggesting an interference with cell biological activity, resulting in less reliable viability data. On the other hand, R...Continue Reading

References

Mar 1, 1972·Toxicology and Applied Pharmacology·L PhillipsW A Akers
Sep 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·J W SangerB M Jockusch
Jun 1, 1995·Burns : Journal of the International Society for Burn Injuries·D J Wainwright
Mar 15, 2000·Journal of Pharmacological and Toxicological Methods·M K RobinsonM A Perkins
May 1, 2001·Bulletin of Experimental Biology and Medicine·E G Kolokol'chikovaV P Tumanov
Nov 27, 2001·Science·E CukiermanK M Yamada
Mar 22, 2002·Advances in Skin & Wound Care·S L HansenC N Paul
Aug 14, 2002·The British Journal of Dermatology·A el-GhalbzouriM Ponec
Dec 4, 2002·Advanced Drug Delivery Reviews·Maria Ponec
Mar 30, 2004·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Thomas WelssKlaus R Schröder
Jul 19, 2005·Clinics in Dermatology·Dorothy M Supp, Steven T Boyce
Oct 19, 2005·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Carine TornierHoward I Maibach
Aug 1, 2006·Regulatory Toxicology and Pharmacology : RTP·Richard A BeckerWilliam T Stott
Feb 13, 2007·European Journal of Cell Biology·Karsten BoehnkeHans-Jürgen Stark
Sep 11, 2007·Burns : Journal of the International Society for Burn Injuries·Clarabelle PhamGuy Maddern
Mar 25, 2008·ALTEX·Thomas Hartung
Dec 15, 2011·Toxicological Sciences : an Official Journal of the Society of Toxicology·Zhiwu Lin, Yvonne Will
Jan 19, 2012·Pharmacology & Therapeutics·Anna AstashkinaDavid W Grainger
Mar 29, 2012·Toxicological Sciences : an Official Journal of the Society of Toxicology·Stephen J Fey, Krzysztof Wrzesinski
Nov 13, 2012·Archives of Toxicology·S MessnerJ M Kelm
Feb 13, 2013·Proceedings of the National Academy of Sciences of the United States of America·Junhee SeokUNKNOWN Inflammation and Host Response to Injury, Large Scale Collaborative Research Program
Mar 19, 2013·Archives of Toxicology·Marcel Leist, Thomas Hartung
Oct 18, 2013·Journal of Laboratory Automation·Markus RimannUrsula Graf-Hausner
Nov 19, 2013·Cytotechnology·Richard Thomas Pace, Karen Jane Leadbetter Burg
Jan 1, 2014·Advanced Drug Delivery Reviews·Adrian Roth, Thomas Singer
Apr 25, 2014·Journal of Biomolecular Screening·Angelika WalzlHelmut Dolznig
Sep 23, 2014·Journal of Biotechnology·Markus RimannUrsula Graf-Hausner

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Citations

Aug 11, 2020·Critical Reviews in Food Science and Nutrition·David Fonseca HernandezLuis Mojica
Jan 20, 2021·Zebrafish·Elizabeth E LeClair
Feb 11, 2021·In Vitro Cellular & Developmental Biology. Animal·Terry Riss, O Joseph Trask
Jun 5, 2021·Tissue Engineering. Part C, Methods·Anthony J DominijanniShay Soker
Jun 3, 2021·International Journal of Molecular Sciences·Andrés TabernillaMathieu Vinken
Nov 19, 2021·Journal of Biomedical Materials Research. Part a·Anna MichopoulouDimitris Bikiaris

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Software Mentioned

GraphPad Prism
Promokine

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