Related Experiment Video
Updated: Nov 6, 2025
Author Spotlight: Insight Into Advances in Prion Diseases Research
Published on: August 11, 2023
The Cell Culture Environment Regulates the Transcription Factor MafB in BV-2 Microglia
Patrick Miller-Rhodes1, Harris A Gelbard1
1Center for Neurotherapeutics Discovery, Department of Neuroscience, Department of Immunology and Microbiology, Department of Neurology, Department of Pediatrics, University of Rochester Medical Center.
Abstract:
Microglia experience dramatic molecular and functional changes when transferred from the central nervous system (CNS) to a cell culture environment. Investigators largely attribute these findings to the loss of CNS-specific microenvironmental cues that dictate the gene-regulatory networks specified by master regulator transcription factors such as V-maf musculoaponeurotic fibrosarcoma oncogene homolog B (MafB). MafB regulates macrophage differentiation and activation by activating or repressing target genes critical to these processes. Here, we show that basal MafB levels in the BV-2 microglial cell line depend on the availability of lipids in the cell culture environment. Depletion of lipids, either by serum deprivation or the use of lipid-depleted serum, reduced MafB protein levels in BV-2 cells. Using live imaging, we also observed the engulfment of apoptotic BV-2 cell debris by neighboring BV-2 cells, highlighting an additional potential source of lipids in the cell culture environment. This observation was supported by experiments showing reduced MafB protein levels in BV-2 cells cultured with various phagocytosis inhibitors (cytochalasin D, annexin V) and reduced BV-2 cell phagocytic activity with serum deprivation. In aggregate, our data suggest that serum exposure regulates the transcription factor MafB in BV-2 cells through direct and indirect mechanisms.
Insights
Lipid availability in cell culture media significantly impacts microglial gene regulation. Serum deprivation reduces the transcription factor V-maf musculoaponeurotic fibrosarcoma oncogene homolog B (MafB) levels in BV-2 cells, affecting their function.
Area of Science:
- Neuroscience
- Cell Biology
- Immunology
Background:
- Microglia, the immune cells of the central nervous system (CNS), undergo significant changes when cultured ex vivo.
- These changes are often attributed to the loss of CNS-specific microenvironmental cues.
- Master regulator transcription factors, like V-maf musculoaponeurotic fibrosarcoma oncogene homolog B (MafB), control microglial gene expression and function.
Purpose of the Study:
- To investigate the role of lipids in regulating the transcription factor MafB in cultured microglial cells.
- To understand the mechanisms by which cell culture environments influence microglial molecular and functional states.
Main Methods:
- Utilized the BV-2 microglial cell line.
- Manipulated lipid availability through serum deprivation and lipid-depleted serum.
- Employed live imaging to observe cellular processes like phagocytosis.
- Assessed MafB protein levels and phagocytic activity.
- Used phagocytosis inhibitors (cytochalasin D, annexin V).
Main Results:
- Basal MafB levels in BV-2 cells were dependent on lipid availability.
- Lipid depletion led to reduced MafB protein levels.
- Observed phagocytosis of apoptotic cell debris by neighboring BV-2 cells, indicating a lipid source.
- Phagocytosis inhibition and serum deprivation reduced MafB levels.
- Serum deprivation decreased BV-2 cell phagocytic activity.
Conclusions:
- Serum exposure regulates the transcription factor MafB in BV-2 cells via direct and indirect pathways.
- Lipid availability is a critical factor influencing microglial phenotype in cell culture.
- Cellular processes like phagocytosis contribute to lipid availability in vitro, impacting gene regulation.
Related Concept Videos
01:30General Transcription Factors
02:23Master Transcription Regulators