RT Journal Article
SR Electronic
T1 Systematic comparison of culture media uncovers phenotypic shift of human microglia defined by reduced reliance to CSF1R signaling
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.07.14.500101
DO 10.1101/2022.07.14.500101
A1 Marie-France Dorion
A1 Moein Yaqubi
A1 Hunter J. Murdoch
A1 Jeffery A. Hall
A1 Roy Dudley
A1 Jack P. Antel
A1 Thomas M. Durcan
A1 Luke M. Healy
YR 2022
UL http://biorxiv.org/content/early/2022/07/14/2022.07.14.500101.abstract
AB Efforts to understand microglia function in health and diseases have been hindered by the lack of culture models that recapitulate in situ cellular properties. In recent years, the use of serum-free media with brain-derived growth factors (CSF1R ligands and TGF-β1/2) have been favored for the maintenance of rodent microglia as they promote morphological features observed in situ. Here we study the functional and transcriptomic impacts of such media on human microglia. Media formulation had little impact on microglia transcriptome assessed by RNA sequencing which was sufficient to significantly alter microglia capacity to phagocytose myelin debris and to elicit an inflammatory response to lipopolysaccharide. When compared to immediately ex vivo microglia from the same donors, the addition of fetal bovine serum to culture media, but not growth factors, was found to aid in the maintenance of key signature genes including those involved in phagocytic processes. A phenotypic shift characterized by CSF1R downregulation in culture correlated with a lack of reliance on CSF1R signaling for survival. Consequently, no improvement in cell survival was observed following culture supplementation with CSF1R ligands. Our study provides better understanding of human microglia in culture, with observations that diverge from those previously made in rodent microglia.Competing Interest StatementThe authors have declared no competing interest.