Anti-inflammatory mechanisms in cancer research : Characterization of a distinct M2-like macrophage model derived from the THP-1 cell line
© 2023 The Authors. Cancer Medicine published by John Wiley & Sons Ltd..
AIMS: Macrophages play an essential role in cancer development. Tumor-associated macrophages (TAMs) have predominantly M2-like attributes that are associated with tumor progression and poor patient survival. Numerous methods have been reported for differentiating and polarizing macrophages in vitro, but there is no standardized and validated model for creating TAMs. Primary cells show varying cytokine responses depending on their origin and functional studies utilizing these cells may lack generalization and validity. A distinct cell line-derived TAM-like M2 subtype is required to investigate the mechanisms mediated by anti-inflammatory TAMs in vitro. Our previous work demonstrated a standardized protocol for creating an M2 subtype derived from a human THP-1 cell line. The cell expression profile, however, has not been validated. The aim of this study was to characterize and validate the TAM-like M2 subtype macrophage created based on our protocol to introduce them as a standardized model for cancer research.
METHODS AND RESULTS: Using qRT-PCR and ELISA, we demonstrated that proinflammatory, anti-inflammatory, and tumor-associated marker expression changed during THP-1-derived marcrophage development in vitro, mimicking a TAM-related profile (e.g., TNFα, IL-1β). The anti-inflammatory marker IL-8/CXCL8, however, is most highly expressed in young M0 macrophages. Flow cytometry showed increased expression of CD206 in the final TAM-like M2 macrophage. Single-cell RNA-sequencing analysis of primary human monocytes and colon cancer tissue macrophages demonstrated that cell line-derived M2 macrophages resembled a TAM-related gene profile.
CONCLUSIONS: The THP-1-derived M2 macrophage based on a standardized cell line model represents a distinct anti-inflammatory TAM-like phenotype with an M2a subtype profile. This model may provide a basis for in vitro investigation of functional mechanisms in a variety of anti-inflammatory settings, particularly colon cancer development.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:12 |
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| Enthalten in: |
Cancer medicine - 12(2023), 23 vom: 19. Dez., Seite 21172-21187 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Scheurlen, Katharina M [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 19.12.2023 Date Revised 23.01.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1002/cam4.6681 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 100 | 1 | |a Scheurlen, Katharina M |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Anti-inflammatory mechanisms in cancer research |b Characterization of a distinct M2-like macrophage model derived from the THP-1 cell line |
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| 500 | |a Date Completed 19.12.2023 | ||
| 500 | |a Date Revised 23.01.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2023 The Authors. Cancer Medicine published by John Wiley & Sons Ltd. | ||
| 520 | |a AIMS: Macrophages play an essential role in cancer development. Tumor-associated macrophages (TAMs) have predominantly M2-like attributes that are associated with tumor progression and poor patient survival. Numerous methods have been reported for differentiating and polarizing macrophages in vitro, but there is no standardized and validated model for creating TAMs. Primary cells show varying cytokine responses depending on their origin and functional studies utilizing these cells may lack generalization and validity. A distinct cell line-derived TAM-like M2 subtype is required to investigate the mechanisms mediated by anti-inflammatory TAMs in vitro. Our previous work demonstrated a standardized protocol for creating an M2 subtype derived from a human THP-1 cell line. The cell expression profile, however, has not been validated. The aim of this study was to characterize and validate the TAM-like M2 subtype macrophage created based on our protocol to introduce them as a standardized model for cancer research | ||
| 520 | |a METHODS AND RESULTS: Using qRT-PCR and ELISA, we demonstrated that proinflammatory, anti-inflammatory, and tumor-associated marker expression changed during THP-1-derived marcrophage development in vitro, mimicking a TAM-related profile (e.g., TNFα, IL-1β). The anti-inflammatory marker IL-8/CXCL8, however, is most highly expressed in young M0 macrophages. Flow cytometry showed increased expression of CD206 in the final TAM-like M2 macrophage. Single-cell RNA-sequencing analysis of primary human monocytes and colon cancer tissue macrophages demonstrated that cell line-derived M2 macrophages resembled a TAM-related gene profile | ||
| 520 | |a CONCLUSIONS: The THP-1-derived M2 macrophage based on a standardized cell line model represents a distinct anti-inflammatory TAM-like phenotype with an M2a subtype profile. This model may provide a basis for in vitro investigation of functional mechanisms in a variety of anti-inflammatory settings, particularly colon cancer development | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a RNA-Seq | |
| 650 | 4 | |a THP-1 cells | |
| 650 | 4 | |a colon cancer | |
| 650 | 4 | |a colonic neoplasms | |
| 650 | 4 | |a immunology | |
| 650 | 4 | |a inflammation | |
| 650 | 4 | |a tumor-associated macrophages | |
| 650 | 7 | |a Anti-Inflammatory Agents |2 NLM | |
| 700 | 1 | |a Snook, Dylan L |e verfasserin |4 aut | |
| 700 | 1 | |a Littlefield, Andrew B |e verfasserin |4 aut | |
| 700 | 1 | |a George, Joan B |e verfasserin |4 aut | |
| 700 | 1 | |a Parks, Mary A |e verfasserin |4 aut | |
| 700 | 1 | |a Beal, Robert J |e verfasserin |4 aut | |
| 700 | 1 | |a MacLeod, Anne |e verfasserin |4 aut | |
| 700 | 1 | |a Riggs, Daniel W |e verfasserin |4 aut | |
| 700 | 1 | |a Gaskins, Jeremy T |e verfasserin |4 aut | |
| 700 | 1 | |a Chariker, Julia |e verfasserin |4 aut | |
| 700 | 1 | |a Rouchka, Eric C |e verfasserin |4 aut | |
| 700 | 1 | |a Galandiuk, Susan |e verfasserin |4 aut | |
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