PT  - JOURNAL ARTICLE
AU  - Marie-France Dorion
AU  - Moein Yaqubi
AU  - Hunter J. Murdoch
AU  - Jeffery A. Hall
AU  - Roy Dudley
AU  - Jack P. Antel
AU  - Thomas M. Durcan
AU  - Luke M. Healy
TI  - Systematic comparison of culture media uncovers phenotypic shift of human microglia defined by reduced reliance to CSF1R signaling
AID  - 10.1101/2022.07.14.500101
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.07.14.500101
4099  - http://biorxiv.org/content/early/2022/07/14/2022.07.14.500101.short
4100  - http://biorxiv.org/content/early/2022/07/14/2022.07.14.500101.full
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.