Comparison of THP-1 Macrophages Viability in Different Types of Culture Vessel: Comparison of Thp-1 Macrophages Viability in Different Types of Culture Vessel

Zahidah Nasuha Mohd Yasin; Fatin Najiah Mohd Idrus; Get Bee Yvonne-Tee

doi:10.11594/jtls.13.02.14

Authors

Zahidah Nasuha Mohd Yasin Biomedicine Programme, School of Health Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia
Fatin Najiah Mohd Idrus Biomedicine Programme, School of Health Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia
Get Bee Yvonne-Tee Biomedicine Programme, School of Health Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia

DOI:

https://doi.org/10.11594/jtls.13.02.14

Keywords:

Apoptosis, culture vessel, Macrophage, THP-1, Viability

Abstract

The ox-LDL generated apoptotic bodies using THP-1 macrophage is a useful tool to study foam cell formation in atherosclerosis. However, the common problem is the cells in the negative control (i.e., absence of ox-LDL) undergo apoptosis. Therefore, the type of cell culture vessel was proposed to be the key factor contributing to cell apoptosis. The THP-1 cells were differentiated into M1 macrophages using 10 ng/μL PMA, 5 ng/μL LPS, and 20 ng/μL IFN-? while 5 ng/μL PMA, 20 ng/μL IL-4 and 20 ng/μL IL-13 were used to differentiate THP-1 into M2 macrophages. Two types of cell culture vessels (6-well plate and T25 flask) were used to culture the macrophages. The cells were subsequently stained using Annexin V-FITC and Propidium Iodide prior to flow cytometry analysis. Interestingly, both M1 and M2 macrophages cultured in the T25 flask resulted in a significantly higher percentage of cell viability compared to macrophages cultured in 6-well plate [M1: 84.15% ± 4.39 vs 8.02% ± 1.55, p < 0.0001; M2: 95.95% ± 1.74 vs 10.50% ± 0.05, p < 0.0001]. In summary, the type of culture vessel is a vital factor in determining cell viability attributed to the surface area and cell seeding density in different types of vessels.

Keywords: Apoptosis, cell viability, culture vessel, macrophage, THP-1

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Comparison of THP-1 Macrophages Viability in Different Types of Culture Vessel