Monitoring of Cell Viability and Proliferation in Hydrogel-Encapsulated System by Resazurin Assay

Xiao, Jing; Zhang, Ying; Wang, Jianzheng; Wu, Yu; Wang, Wei; Ma, Xiaojun

doi:10.1007/s12010-010-8975-3

Cited by 48 publications

(38 citation statements)

References 18 publications

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“…CTB at 10% correspond to 44 μM as demonstrated by Xiao et al . [30]. Fluorescence intensity of resazurin solution was measured with the GloMax ® -Multi Microplate Multimode Reader (Promega Corporation) by using the optical kit of filters for resorufin, typically used for CTB assay having a preset excitation peak wavelength at 525 nm and an emission wavelength in the range of 580–640 nm.…”

Section: Materials and Measurement Methodsmentioning

confidence: 99%

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: Materials and Measurement Methodsmentioning

confidence: 99%

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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“…Therefore we chose to work with the open-source software ImageJ and developed an in-house automated macro for spheroid analysis to facilitate image analysis within the scientific community. Apart from volume, cell viability within the spheroid can be assessed using metabolic assays like the reduction of Resazurin [24] or measuring ATP [19]. These assays are convenient and quick however they have not been properly validated yet for use in 3D cultures.…”

Section: Introductionmentioning

confidence: 99%

Multiplexing Spheroid Volume, Resazurin and Acid Phosphatase Viability Assays for High-Throughput Screening of Tumour Spheroids and Stem Cell Neurospheres

et al. 2014

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Three-dimensional cell culture has many advantages over monolayer cultures, and spheroids have been hailed as the best current representation of small avascular tumours in vitro. However their adoption in regular screening programs has been hindered by uneven culture growth, poor reproducibility and lack of high-throughput analysis methods for 3D. The objective of this study was to develop a method for a quick and reliable anticancer drug screen in 3D for tumour and human foetal brain tissue in order to investigate drug effectiveness and selective cytotoxic effects. Commercially available ultra-low attachment 96-well round-bottom plates were employed to culture spheroids in a rapid, reproducible manner amenable to automation. A set of three mechanistically different methods for spheroid health assessment (Spheroid volume, metabolic activity and acid phosphatase enzyme activity) were validated against cell numbers in healthy and drug-treated spheroids. An automated open-source ImageJ macro was developed to enable high-throughput volume measurements. Although spheroid volume determination was superior to the other assays, multiplexing it with resazurin reduction and phosphatase activity produced a richer picture of spheroid condition. The ability to distinguish between effects on malignant and the proliferating component of normal brain was tested using etoposide on UW228-3 medulloblastoma cell line and human neural stem cells. At levels below 10 µM etoposide exhibited higher toxicity towards proliferating stem cells, whereas at concentrations above 10 µM the tumour spheroids were affected to a greater extent. The high-throughput assay procedures use ready-made plates, open-source software and are compatible with standard plate readers, therefore offering high predictive power with substantial savings in time and money.

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“…Cell activity was measured by resazurin assay that measures the aerobic respiration of the cells to assess cell viability [33]. Nucleus pulposus (NP) tissues and Annulus fibrosus (AF) tissues from each disc were incubated in medium with 50μM resazurin sodium salt (Sigma-Aldrich, Buchs Switzerland) for five hours.…”

Section: Methodsmentioning

confidence: 99%

Loading-Induced Heat-Shock Response in Bovine Intervertebral Disc Organ Culture

et al. 2016

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Mechanical loading has been shown to affect cell viability and matrix maintenance in the intervertebral disc (IVD) but there is no investigation on how cells survive mechanical stress and whether the IVD cells perceive mechanical loading as stress and respond by expression of heat shock proteins. This study investigates the stress response in the IVD in response to compressive loading. Bovine caudal disc organ culture was used to study the effect of physiological range static loading and dynamic loading. Cell activity, gene expression and immunofluorescence staining were used to analyze the cell response. Cell activity and cytoskeleton of the cells did not change significantly after loading. In gene expression analysis, significant up-regulation of heat shock protein-70 (HSP70) was observed in nucleus pulposus after two hours of loading. However, the expression of the matrix remodeling genes did not change significantly after loading. Similarly, expressions of stress response and matrix remodeling genes changed with application and removal of the dynamic loading. The results suggest that stress response was induced by physiological range loading without significantly changing cell activity and upregulating matrix remodeling. This study provides direct evidence on loading induced stress response in IVD cells and contributes to our understanding in the mechanoregulation of intervertebral disc cells.

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Monitoring of Cell Viability and Proliferation in Hydrogel-Encapsulated System by Resazurin Assay

Cited by 48 publications

References 18 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

Multiplexing Spheroid Volume, Resazurin and Acid Phosphatase Viability Assays for High-Throughput Screening of Tumour Spheroids and Stem Cell Neurospheres

Loading-Induced Heat-Shock Response in Bovine Intervertebral Disc Organ Culture

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