THP-1巨噬细胞在不同类型培养容器中生存能力的比较

Q4 Agricultural and Biological Sciences Journal of Tropical Life Science Pub Date : 2023-05-25 DOI:10.11594/jtls.13.02.14
Z. Yasin, Fatin Idrus, G. Tee
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引用次数: 0

摘要

利用THP-1巨噬细胞生成ox-LDL产生的凋亡小体是研究动脉粥样硬化中泡沫细胞形成的有用工具。然而,常见的问题是阴性对照(即缺乏ox-LDL)的细胞发生凋亡。因此,细胞培养容器的类型被认为是导致细胞凋亡的关键因素。以10 ng/μL PMA、5 ng/μL LPS和20 ng/μL IFN-?5 ng/μL PMA、20 ng/μL IL-4和20 ng/μL IL-13诱导THP-1向M2巨噬细胞分化。采用两种细胞培养容器(6孔板和T25烧瓶)培养巨噬细胞。流式细胞术分析前,用Annexin V-FITC和碘化丙啶对细胞进行染色。有趣的是,在T25烧瓶中培养的M1和M2巨噬细胞的细胞存活率明显高于在6孔板中培养的巨噬细胞[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]。总之,培养容器的类型是决定细胞存活率的重要因素,因为不同类型的容器的表面积和细胞播种密度。
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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 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.
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来源期刊
Journal of Tropical Life Science
Journal of Tropical Life Science Environmental Science-Ecology
CiteScore
1.00
自引率
0.00%
发文量
46
审稿时长
12 weeks
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