IMDM-20 enhances neutrophilic features during 1.3 % DMSO-mediated differentiation of HL-60 cells
The promyelocytic HL-60 cell line can be differentiated toward neutrophil-like cells and has been historically used as a surrogate to study human neutrophil biology in vitro. Multiple differentiation protocols have been reported to generate neutrophil-like HL-60 cells, with limited consideration of...
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Elsevier
2025-09-01
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| Series: | Biochemistry and Biophysics Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580825003024 |
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| author | Jorge Andrés Cázares-Preciado José Antonio Cruz-Cárdenas Alejandra López-Arredondo Marco V. Gallardo-Camarena Marion E.G. Brunck |
| author_facet | Jorge Andrés Cázares-Preciado José Antonio Cruz-Cárdenas Alejandra López-Arredondo Marco V. Gallardo-Camarena Marion E.G. Brunck |
| author_sort | Jorge Andrés Cázares-Preciado |
| collection | DOAJ |
| description | The promyelocytic HL-60 cell line can be differentiated toward neutrophil-like cells and has been historically used as a surrogate to study human neutrophil biology in vitro. Multiple differentiation protocols have been reported to generate neutrophil-like HL-60 cells, with limited consideration of how methodological variations might influence cell identity and functions. Here, we conducted a systematic search of the PubMed database, to investigate the current heterogeneity in published protocols used to differentiate HL-60 towards neutrophil-like cells. Research articles published in English between January 9th, 2020, and January 9th, 2025, were identified using the key words “neutrophil-like cell” and “HL-60”. Metadata of included research papers were charted and analyzed to evaluate the most reported HL-60 cells culture protocols. A total of 71 studies published in 5 years employed 41 distinct protocols. The 3 most prevalent conditions to maintain HL-60 cells were IMDM with 20 % FBS (IMDM-20), DMEM with 10 % FBS (DMEM-10), and RPMI-1640 with 10 % FBS (RPMI-10). Over 90 % of protocols applied 1–1.57 % DMSO as differentiating agent to produce neutrophil-like cells. In the laboratory, we compared the 3 most common culture media applied during neutrophil-like cell differentiation with 1.3 % DMSO over 5 and 7 days. Using IMDM-20 led to the highest proliferation rate and cell yield during differentiation. Neutrophil-like cells produced in IMDM-20 and RPMI-10 exhibited significantly higher proportions of CD15+CD11b+ cells, and significantly higher bacterial clearance compared to DMEM-10. Culture media did not affect phagocytosis, but using RPMI-10 over 5 days led to significantly higher ability to produce ROS. IMDM-20 produced significantly more IL-6 and IL-1β in culture supernatant following stimulation with opsonized E. coli. Overall, the results support the use of IMDM-20 with 1.3 % DMSO to differentiate HL-60 to study neutrophil biology in vitro. |
| format | Article |
| id | doaj-art-9a04b4701b1a47cd99e4005efe72a90f |
| institution | Kabale University |
| issn | 2405-5808 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
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| series | Biochemistry and Biophysics Reports |
| spelling | doaj-art-9a04b4701b1a47cd99e4005efe72a90f2025-08-26T04:14:26ZengElsevierBiochemistry and Biophysics Reports2405-58082025-09-014310221510.1016/j.bbrep.2025.102215IMDM-20 enhances neutrophilic features during 1.3 % DMSO-mediated differentiation of HL-60 cellsJorge Andrés Cázares-Preciado0José Antonio Cruz-Cárdenas1Alejandra López-Arredondo2Marco V. Gallardo-Camarena3Marion E.G. Brunck4Tecnológico de Monterrey, Escuela de Ingeniería y Ciencias, Av. Eugenio Garza Sada 2501 Sur, Tecnologico, 64849, Monterrey, Nuevo León, MexicoTecnológico de Monterrey, Escuela de Ingeniería y Ciencias, Av. Eugenio Garza Sada 2501 Sur, Tecnologico, 64849, Monterrey, Nuevo León, MexicoTecnológico de Monterrey, Escuela de Ingeniería y Ciencias, Av. Eugenio Garza Sada 2501 Sur, Tecnologico, 64849, Monterrey, Nuevo León, MexicoTecnológico de Monterrey, Escuela de Ingeniería y Ciencias, Av. Eugenio Garza Sada 2501 Sur, Tecnologico, 64849, Monterrey, Nuevo León, MexicoDepartamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Apdo. Postal 70228, Ciudad Universitaria, Ciudad de México, 04510, Mexico; Tecnologico de Monterrey, Institute for Obesity Research, Av. Eugenio Garza Sada 2501 Sur, C.P., 64849, Monterrey, Nuevo León, Mexico; Corresponding author. Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Apdo. Postal 70228, Ciudad Universitaria, Ciudad de México, 04510, Mexico.The promyelocytic HL-60 cell line can be differentiated toward neutrophil-like cells and has been historically used as a surrogate to study human neutrophil biology in vitro. Multiple differentiation protocols have been reported to generate neutrophil-like HL-60 cells, with limited consideration of how methodological variations might influence cell identity and functions. Here, we conducted a systematic search of the PubMed database, to investigate the current heterogeneity in published protocols used to differentiate HL-60 towards neutrophil-like cells. Research articles published in English between January 9th, 2020, and January 9th, 2025, were identified using the key words “neutrophil-like cell” and “HL-60”. Metadata of included research papers were charted and analyzed to evaluate the most reported HL-60 cells culture protocols. A total of 71 studies published in 5 years employed 41 distinct protocols. The 3 most prevalent conditions to maintain HL-60 cells were IMDM with 20 % FBS (IMDM-20), DMEM with 10 % FBS (DMEM-10), and RPMI-1640 with 10 % FBS (RPMI-10). Over 90 % of protocols applied 1–1.57 % DMSO as differentiating agent to produce neutrophil-like cells. In the laboratory, we compared the 3 most common culture media applied during neutrophil-like cell differentiation with 1.3 % DMSO over 5 and 7 days. Using IMDM-20 led to the highest proliferation rate and cell yield during differentiation. Neutrophil-like cells produced in IMDM-20 and RPMI-10 exhibited significantly higher proportions of CD15+CD11b+ cells, and significantly higher bacterial clearance compared to DMEM-10. Culture media did not affect phagocytosis, but using RPMI-10 over 5 days led to significantly higher ability to produce ROS. IMDM-20 produced significantly more IL-6 and IL-1β in culture supernatant following stimulation with opsonized E. coli. Overall, the results support the use of IMDM-20 with 1.3 % DMSO to differentiate HL-60 to study neutrophil biology in vitro.http://www.sciencedirect.com/science/article/pii/S2405580825003024HL-60NeutrophilsCulture mediaDifferentiationNeutrophil-like cells |
| spellingShingle | Jorge Andrés Cázares-Preciado José Antonio Cruz-Cárdenas Alejandra López-Arredondo Marco V. Gallardo-Camarena Marion E.G. Brunck IMDM-20 enhances neutrophilic features during 1.3 % DMSO-mediated differentiation of HL-60 cells Biochemistry and Biophysics Reports HL-60 Neutrophils Culture media Differentiation Neutrophil-like cells |
| title | IMDM-20 enhances neutrophilic features during 1.3 % DMSO-mediated differentiation of HL-60 cells |
| title_full | IMDM-20 enhances neutrophilic features during 1.3 % DMSO-mediated differentiation of HL-60 cells |
| title_fullStr | IMDM-20 enhances neutrophilic features during 1.3 % DMSO-mediated differentiation of HL-60 cells |
| title_full_unstemmed | IMDM-20 enhances neutrophilic features during 1.3 % DMSO-mediated differentiation of HL-60 cells |
| title_short | IMDM-20 enhances neutrophilic features during 1.3 % DMSO-mediated differentiation of HL-60 cells |
| title_sort | imdm 20 enhances neutrophilic features during 1 3 dmso mediated differentiation of hl 60 cells |
| topic | HL-60 Neutrophils Culture media Differentiation Neutrophil-like cells |
| url | http://www.sciencedirect.com/science/article/pii/S2405580825003024 |
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