Functional and Metabolic Characterization of Microglia Culture in a Defined Medium.

Alejandro Montilla,Carlos Matute,Alazne Zabala,María Domercq

doi:10.3389/fncel.2020.00022

Abstract

Microglia are the endogenous immune cells of the brain and act as sensor of infection and pathologic injury to the brain, leading to a rapid plastic process of activation that culminates in the endocytosis and phagocytosis of damaged tissue. Microglia cells are the most plastic cells in the brain. Microglia isolation from their environment as well as culturing them in the presence of serum alter their function and lead to a rapid loss of their signature gene expression. Previous studies have identified pivotal factors allowing microglia culture in the absence of serum. Here, we have further characterized the function, expression of markers, metabolic status and response to pro and anti-inflammatory stimulus of microglia isolated by magnetic-activated cell sorting and cultured in a chemically defined medium. We have compared this new method with previous traditional protocols of culturing microglia that use high concentrations of serum.

Highlights

Microglial cells are the resident macrophages of the central nervous system (CNS)
Morphological analysis based on Iba1 immunostaining showed a significant increase in the number of processes and a decrease in the cell body area in microglia cultured in defined medium (Figure 1)
Microglia acquired a highly ramified morphology, with multiple processes branching off the soma, resembling the typical microglia morphology presented in physiological conditions in the brain

Summary

Introduction

Microglial cells are the resident macrophages of the central nervous system (CNS) They arise from early myeloid progenitors in the embryonic yolk sac that migrate and colonize CNS (Ginhoux et al, 2010; Kierdorf et al, 2013). Microglia are characterized by a continuous surveillance of their environment, in order to maintain tissue integrity and homeostasis (Davalos et al, 2005; Nimmerjahn et al, 2005). They play a pivotal role in a wide and heterogeneous range of physiological responses, actively interacting with almost all CNS cell types. Microglia is involved in maintaining the homeostasis of the baseline neurogenic cascade through the elimination of neural precursor cells (NPCs; Sierra et al, 2010)

Methods

Results

Discussion

Conclusion