Efficiency of MTT and Trypan Blue Assays for Detection of Viability and Recovery of Different Frozen Cell Lines

Azzoz, Mohamed Fahmy; Zaghloul, Mostafa Amin; Sayed, Maha Magdy; Soliman, Heba Mohamed; Mounir, Nermin Mohamed; Abdel Shakour, Nehal Kamal; El-Dabae, Wahid Hussein; Aly, Saleh Esmate

doi:10.21608/ejvs.2024.260687.1764

Efficiency of MTT and Trypan Blue Assays for Detection of Viability and Recovery of Different Frozen Cell Lines

Document Type : Original Article

Authors

¹ 8Central Laboratory for Evaluation of Veterinary Biologics (CLEVB), Agricultural Research Center (ARC), Cairo, Egypt

² Central Laboratory for Evaluation of Veterinary Biologics (CLEVB), Agricultural Research Center (ARC), Cairo, Egypt

³ 6Veterinary Serum and Vaccine Research Institute (VSVRI), Agricultural Research Center (ARC), Cairo, Egypt.

⁴ National research center veterinary research division microbiology and immunology department.

⁵ Central Laboratory for Evaluation of Veterinary Biologics (CLEVB), Agricultural Research Center (ARC), Cairo, Egypt.

10.21608/ejvs.2024.260687.1764

Abstract

The quantity of healthy cells in a sample is known as cell viability. For all types of cell culture, measuring cell viability is crucial to ensuring high-quality cell lines through setup, preparation and storage. Although cryopreservation is used in both scientific and clinical research, there are still certain restrictions. At low temperatures, such as −196 °C cells metabolize almost nothing, which has unavoidable side effects, such as genetic drift toward biological variations of cell-associated changes in lipids. Also, cells can be harmed by cryoprotective agents (CPAs), especially when administered in high concentrations So, it is important to use a reliable technique to follow up changes affecting cell viability after cryopreservation which were compared with control freshly subcultured cells. In the present study, the MTT assay was used to correlate cell behavior according to the number of healthy and viable cells. Different kinds of cell cultures were cryopreserved with dimethyl sulfoxide at 10% for variable years in a liquid nitrogen tank . Then, different samples were tested for viability using MTT assay and counted by the trypan blue assay for comparing both results accuracy and confirmed by visual examination in sub-cultured media twenty four hours post-thawing. The MTT assay was easy, fast and showed more accurate results than the trypan blue assay. The visual examination twenty four hours post-subculture indicated that MTT assay can differentiate between healthy and apparently viable cells which have lost their function while the trypan blue assay failed to detect healthy cells in some samples.

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