Resazurin Dye as a Reliable Tool for Determination of Cell Number and Viability in Mesenchymal Stem Cell Culture

Dienstknecht, Thomas; Ehehalt, Katharina; Jenei-Lanzl, Zsuzsa; Zellner, Johannes; Müller, Michael; Berner, Arne; Nerlich, Michael; Angele, Peter

doi:10.1007/s10517-010-1092-z

Cited by 8 publications

(5 citation statements)

References 11 publications

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“…Also Dienstknecht et al . [40] showed similar detection limit for resazurin in 2D cell cultures (500 cells). The lower detection limit in 3D cell cultures could be due to the high porosity: pores retain a small amount of resazurin which is not analyzed for fluorescence which is underestimated.…”

Section: Discussionmentioning

confidence: 82%

A First Approach to Evaluate the Cell Dose in Highly Porous Scaffolds By Using a Nondestructive Metabolic Method

Divieto

Sassi

2015

Future Sci. OA

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Background:In cell-based therapies, in vitro studies on biomimetic cell–scaffold constructs can facilitate the determination of the cell dose, a key factor in guaranteeing the effectiveness of the treatment. However, highly porous scaffolds do not allow a nondestructive evaluation of the cell number. Our objective was to develop a nondestructive method for human mesenchymal stem cells dose evaluation in a highly porous scaffold for bone regeneration.Materials & measurement method:Proliferation trend of human mesenchymal stem cells on Biocoral® scaffolds was measured by a resazurin-based assay here optimized for 3D cultures. The method allows to noninvasively follow the cell proliferation on biocorals over 3 weeks with very high reproducibility.Conclusion:This reliable method could be a powerful tool in cell-based therapies for cell dose determination.

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Section: Discussionmentioning

confidence: 82%

A First Approach to Evaluate the Cell Dose in Highly Porous Scaffolds By Using a Nondestructive Metabolic Method

Divieto

Sassi

2015

Future Sci. OA

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“…To further confirm and examine whether the observed enhancing/inhibitory effect occurred only during early treatment or throughout the duration, cells pretreated for 0 (0D-PT), 9 (9D-PT), 12 (12D-PT) or 30 (30D-PT) days of resveratrol or BM were plated at equal low density and treatment was continued until cells of one treatment group reached 100% confluency (regardless of the confluency of the other groups), when resazurin assay was carried out to assess the relative number of live cells in each treatment condition. The OD reading of resazurin solution is inversely correlated to the numbers of live cells [74]. At all 4 different pretreatment time points, 0.1 µM resveratrol treatment resulted in the lowest OD reading and therefore the highest cell numbers, whereas higher concentrations of resveratrol resulted in a dosage dependent increase in OD readings (Figure 1.B.…”

Section: Resultsmentioning

confidence: 91%

Resveratrol Exerts Dosage and Duration Dependent Effect on Human Mesenchymal Stem Cell Development

et al. 2012

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Studies in the past have illuminated the potential benefit of resveratrol as an anticancer (pro-apoptosis) and life-extending (pro-survival) compound. However, these two different effects were observed at different concentration ranges. Studies of resveratrol in a wide range of concentrations on the same cell type are lacking, which is necessary to comprehend its diverse and sometimes contradictory cellular effects. In this study, we examined the effects of resveratrol on cell self-renewal and differentiation of human mesenchymal stem cells (hMSCs), a type of adult stem cells that reside in a number of tissues, at concentrations ranging from 0.1 to 10 µM after both short- and long-term exposure. Our results reveal that at 0.1 µM, resveratrol promotes cell self-renewal by inhibiting cellular senescence, whereas at 5 µM or above, resveratrol inhibits cell self-renewal by increasing senescence rate, cell doubling time and S-phase cell cycle arrest. At 1 µM, its effect on cell self-renewal is minimal but after long-term exposure it exerts an inhibitory effect, accompanied with increased senescence rate. At all concentrations, resveratrol promotes osteogenic differentiation in a dosage dependent manner, which is offset by its inhibitory effect on cell self-renewal at high concentrations. On the contrary, resveratrol suppresses adipogenic differentiation during short-term exposure but promotes this process after long-term exposure. Our study implicates that resveratrol is the most beneficial to stem cell development at 0.1 µM and caution should be taken in applying resveratrol as an anticancer therapeutic agent or nutraceutical supplement due to its dosage dependent effect on hMSCs.

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“…Cell viability was assessed at day 2, 4, 7, 10, 14, 18 and day 21. Cell viability is not affected by this assay [39]. …”

Section: Methodsmentioning

confidence: 99%

Higher Ratios of Hyaluronic Acid Enhance Chondrogenic Differentiation of Human MSCs in a Hyaluronic Acid–Gelatin Composite Scaffold

Pfeifer¹,

Berner²,

Koch³

et al. 2016

Materials

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Mesenchymal stem cells (MSCs) seeded on specific carrier materials are a promising source for the repair of traumatic cartilage injuries. The best supportive carrier material has not yet been determined. As natural components of cartilage’s extracellular matrix, hyaluronic acid and collagen are the focus of biomaterial research. In order to optimize chondrogenic support, we investigated three different scaffold compositions of a hyaluronic acid (HA)-gelatin based biomaterial. Methods: Human MSCs (hMSCs) were seeded under vacuum on composite scaffolds of three different HA-gelatin ratios and cultured in chondrogenic medium for 21 days. Cell-scaffold constructs were assessed at different time points for cell viability, gene expression patterns, production of cartilage-specific extracellular matrix (ECM) and for (immuno-)histological appearance. The intrinsic transforming growth factor beta (TGF-beta) uptake of empty scaffolds was evaluated by determination of the TGF-beta concentrations in the medium over time. Results: No significant differences were found for cell seeding densities and cell viability. hMSCs seeded on scaffolds with higher ratios of HA showed better cartilage-like differentiation in all evaluated parameters. TGF-beta uptake did not differ between empty scaffolds. Conclusion: Higher ratios of HA support the chondrogenic differentiation of hMSCs seeded on a HA-gelatin composite scaffold.

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Resazurin Dye as a Reliable Tool for Determination of Cell Number and Viability in Mesenchymal Stem Cell Culture

Cited by 8 publications

References 11 publications

A First Approach to Evaluate the Cell Dose in Highly Porous Scaffolds By Using a Nondestructive Metabolic Method

A First Approach to Evaluate the Cell Dose in Highly Porous Scaffolds By Using a Nondestructive Metabolic Method

Resveratrol Exerts Dosage and Duration Dependent Effect on Human Mesenchymal Stem Cell Development

Higher Ratios of Hyaluronic Acid Enhance Chondrogenic Differentiation of Human MSCs in a Hyaluronic Acid–Gelatin Composite Scaffold

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