A new perspective reexploration of neonatal mice glial culture in vitro and inflammatory model / 中国药理学通报

ABSTRACT

Aim To revisit the long-established murine glial cul-ture-base inflammation model, in order to explore the possible improvements of current methods. Methods The proportion and purity of glial cells were characterized by qPCR, flow cytometr) and immunofluorescence. After stimulation with LPS, their inflammation response was evaluated at both mRNA and protein levels. Results Mixed glial cells were stimulated by LPS (IOC jig L-1) , and the expression of inflammatory factors increased more significantly than that of microglial. For example, at PDL-coated condition, TNF-a increased more significantly in mixec glial cells ( 903. 8 ± 322. 2 ) ng L-1 than that in microglia (565.4 ± 159. 8)ng • L-1. When cultured as a glial mixture, cells grew better on a matrigel-coated surface. However, when cultured in uncoated condition, purified microglia were more sensitive to LPS [TNF-ot; (6861.4 ± 1606.6) ng L-1[. Conclusions Matrigel, as a newly emerged coating material, is proved to be better than the conventional PDL-coating for cultu-ring mixed glial cells, which improves cell viability and sensitivity. Multiplex inflammatory factor assays should be used for quantifying inflammatory response, rather than relying on qPCR alone.