Comparison of THP-1 Macrophages Viability in Different Types of Culture Vessel
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 t...
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| Format: | Article |
| Language: | English |
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University of Brawijaya
2023-06-01
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| Series: | Journal of Tropical Life Science |
| Subjects: | |
| Online Access: | https://jtrolis.ub.ac.id/index.php/jtrolis/article/view/2507 |
| _version_ | 1797688451605200896 |
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| author | Zahidah Nasuha Mohd Yasin Fatin Najiah Mohd Idrus Get Bee Yvonne-Tee |
| author_facet | Zahidah Nasuha Mohd Yasin Fatin Najiah Mohd Idrus Get Bee Yvonne-Tee |
| author_sort | Zahidah Nasuha Mohd Yasin |
| collection | DOAJ |
| description | 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 |
| first_indexed | 2024-03-12T01:31:11Z |
| format | Article |
| id | doaj.art-5bb921329fbe40f9a1c20c386269a4f1 |
| institution | Directory Open Access Journal |
| issn | 2087-5517 2527-4376 |
| language | English |
| last_indexed | 2024-03-12T01:31:11Z |
| publishDate | 2023-06-01 |
| publisher | University of Brawijaya |
| record_format | Article |
| series | Journal of Tropical Life Science |
| spelling | doaj.art-5bb921329fbe40f9a1c20c386269a4f12023-09-12T01:32:28ZengUniversity of BrawijayaJournal of Tropical Life Science2087-55172527-43762023-06-0113235936810.11594/jtls.13.02.142452Comparison of THP-1 Macrophages Viability in Different Types of Culture VesselZahidah Nasuha Mohd Yasin0Fatin Najiah Mohd Idrus1Get Bee Yvonne-Tee2Biomedicine Programme, School of Health Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, MalaysiaBiomedicine Programme, School of Health Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, MalaysiaBiomedicine Programme, School of Health Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, MalaysiaThe 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-1https://jtrolis.ub.ac.id/index.php/jtrolis/article/view/2507apoptosisculture vesselmacrophagethp-1viability |
| spellingShingle | Zahidah Nasuha Mohd Yasin Fatin Najiah Mohd Idrus Get Bee Yvonne-Tee Comparison of THP-1 Macrophages Viability in Different Types of Culture Vessel Journal of Tropical Life Science apoptosis culture vessel macrophage thp-1 viability |
| title | Comparison of THP-1 Macrophages Viability in Different Types of Culture Vessel |
| title_full | Comparison of THP-1 Macrophages Viability in Different Types of Culture Vessel |
| title_fullStr | Comparison of THP-1 Macrophages Viability in Different Types of Culture Vessel |
| title_full_unstemmed | Comparison of THP-1 Macrophages Viability in Different Types of Culture Vessel |
| title_short | Comparison of THP-1 Macrophages Viability in Different Types of Culture Vessel |
| title_sort | comparison of thp 1 macrophages viability in different types of culture vessel |
| topic | apoptosis culture vessel macrophage thp-1 viability |
| url | https://jtrolis.ub.ac.id/index.php/jtrolis/article/view/2507 |
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