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A High-throughput End-point Assay for Viable Mammalian Cell Estimation.

Xianghui Gong¹, Qiangyi Fang, Xuesen Li

¹Graduate School of the Chinese Academy of Sciences, Shanghai, China.

|November 13, 2008

Summary

A new resazurin assay accurately estimates Chinese hamster ovary (CHO) and hybridoma cell concentrations. This high-throughput method is robust to pH and temperature variations, aiding cell culture optimization.

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A Strategy to Identify Compounds that Affect Cell Growth and Survival in Cultured Mammalian Cells at Low-to-Moderate Throughput

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Published on: September 22, 2019

Area of Science:

Biotechnology
Cell Biology
Assay Development

Background:

Accurate cell concentration determination is crucial for bioprocesses.
Existing methods may be time-consuming or resource-intensive.
High-throughput screening demands efficient cell quantification techniques.

Purpose of the Study:

To develop and validate a novel, high-throughput microplate-based assay for quantifying Chinese hamster ovary (CHO) and hybridoma cell concentrations.
To assess the assay's robustness against common cell culture variations.
To establish the assay's utility in optimizing cell culture conditions.

Main Methods:

Development of a single-wavelength colorimetric assay utilizing the non-cytotoxic dye resazurin.
Testing the assay's performance with Chinese hamster ovary (CHO) and hybridoma cell lines.
Evaluation of assay tolerance to variations in pH and temperature.
Determination of viable cell concentration range using a microplate reader at 605 nm.

Main Results:

The resazurin assay effectively estimated viable cell concentrations in the range of 10(5)-10(7) cells/mL.
The assay demonstrated robustness, tolerating pH and temperature fluctuations inherent in cell cultivation and assay procedures.
The method proved suitable for high-throughput screening, enabling rapid analysis of numerous cell culture samples.

Conclusions:

A reliable and efficient resazurin-based microplate assay for viable cell concentration estimation has been established.
The assay's tolerance to environmental variations and high-throughput capability make it valuable for biopharmaceutical research.
This method holds potential for applications in cell culture medium formulation and optimization, particularly within high-throughput screening platforms.