Validation of cell density and viability assays using Cedex automated cell counter

Huang, Li‐Chun; Lin, Wendy; Yagami, Machiko; Tseng, Daphne ChingYu; Miyashita‐Lin, Emily; Singh, Nitasha; Lin, Andy; Shih, Shian-Jiun

doi:10.1016/j.biologicals.2010.01.009

Cited by 21 publications

(23 citation statements)

References 8 publications

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“…For this purpose we standardized the sample preparation, and adjusted the settings in order to obtain viable cell-counts using CHO-K1 and U937 cells that are employed for the biosynthesis and bioassays of recombinant proteins respectively. The suitability of these methodologies was evaluated through a validation according to the ICH Q2R1 guideline [2], [3].…”

Section: Introductionmentioning

confidence: 99%

Validation of three viable-cell counting methods: Manual, semi-automated, and automated

Cadena-Herrera

Lara²,

Ramírez-Ibañez³

et al. 2015

Biotechnology Reports

168

109

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Section: Introductionmentioning

confidence: 99%

Validation of three viable-cell counting methods: Manual, semi-automated, and automated

Cadena-Herrera

Lara²,

Ramírez-Ibañez³

et al. 2015

Biotechnology Reports

168

109

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show abstract

“…This was solved by incorporating a Cedex Cell Counter (Roche, Innovatis), a validated automated cell counting system [25], under the control of the Freedom Evoware software. The Cedex Cell Counter functions with the principle of Trypan blue exclusion staining and records microscopic pictures that are analyzed by an inherent imaging software.…”

Section: Resultsmentioning

confidence: 99%

An Automated HIV-1 Env-Pseudotyped Virus Production for Global HIV Vaccine Trials

et al. 2012

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BackgroundInfections with HIV still represent a major human health problem worldwide and a vaccine is the only long-term option to fight efficiently against this virus. Standardized assessments of HIV-specific immune responses in vaccine trials are essential for prioritizing vaccine candidates in preclinical and clinical stages of development. With respect to neutralizing antibodies, assays with HIV-1 Env-pseudotyped viruses are a high priority. To cover the increasing demands of HIV pseudoviruses, a complete cell culture and transfection automation system has been developed.Methodology/Principal FindingsThe automation system for HIV pseudovirus production comprises a modified Tecan-based Cellerity system. It covers an area of 5×3 meters and includes a robot platform, a cell counting machine, a CO2 incubator for cell cultivation and a media refrigerator. The processes for cell handling, transfection and pseudovirus production have been implemented according to manual standard operating procedures and are controlled and scheduled autonomously by the system. The system is housed in a biosafety level II cabinet that guarantees protection of personnel, environment and the product. HIV pseudovirus stocks in a scale from 140 ml to 1000 ml have been produced on the automated system. Parallel manual production of HIV pseudoviruses and comparisons (bridging assays) confirmed that the automated produced pseudoviruses were of equivalent quality as those produced manually. In addition, the automated method was fully validated according to Good Clinical Laboratory Practice (GCLP) guidelines, including the validation parameters accuracy, precision, robustness and specificity.ConclusionsAn automated HIV pseudovirus production system has been successfully established. It allows the high quality production of HIV pseudoviruses under GCLP conditions. In its present form, the installed module enables the production of 1000 ml of virus-containing cell culture supernatant per week. Thus, this novel automation facilitates standardized large-scale productions of HIV pseudoviruses for ongoing and upcoming HIV vaccine trials.

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“…A typical cell counting procedure for adherent cells involves harvesting adherent cells, removing cells from culture dish, resuspension after centrifugation, staining, and cell counting using high resolution microscopy with drug treatment if required. In addition, the time required to process numerous samples can limit the accuracy and versatility of the procedure [1][2][3]. The retinal pigment epithelium (RPE) cells are a representative model for adherence because they grow in monolayers.…”

Section: Introductionmentioning

confidence: 99%

In Vitro Monitoring of a Cultured Human Retinal Pigment Epithelium Using 1375-nm Spectral-Domain Optical Coherence Tomography

Kim

Togloom

Han

et al. 2017

J. Lightwave Technol.

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Measurement of cell number is important in cell-based in vitro screenings for therapeutic drugs and tissue engineering such as cell culture maintenance, cell plating, and cell growth, as well as monitoring of cell viability, proliferation, and cytotoxicity. We performed cell counting using intensity-based in vitro spectral domain optical coherence tomography (SD-OCT). Adult retinal pigment epithelium cells were cultured on a glass dish to prevent diffuse reflection from the surface, and the dish was tilted by 15° during OCT imaging to reduce coherence noise from specular reflection. Rasterized scanning was performed to generate a 3D volume image with an en face image of the retinal pigment epithelium cell layers. Cell counting was achieved by measuring the density of bright spots after layer extraction and thresholding. Two other cell counting methods were also performed for the purpose of comparison, one using a hemocytometer and the other using water-soluble tetrazolium-1 (WST-1); cell counting by in vitro OCT yielded results better than those from the hemocytometer. Our results showed that in vitro OCT systems can be a powerful tool for estimating and analyzing cell density in a cultured sample without the need for dyeing the sample or causing cell death.

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Validation of cell density and viability assays using Cedex automated cell counter

Cited by 21 publications

References 8 publications

Validation of three viable-cell counting methods: Manual, semi-automated, and automated

Validation of three viable-cell counting methods: Manual, semi-automated, and automated

An Automated HIV-1 Env-Pseudotyped Virus Production for Global HIV Vaccine Trials

In Vitro Monitoring of a Cultured Human Retinal Pigment Epithelium Using 1375-nm Spectral-Domain Optical Coherence Tomography

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