Mgso4

Primary glial cells, mainly comprised of astrocytes and microglia, were obtained as previously described (36 ). Briefly, we isolated the cerebrum from postnatal day 2 mice of either sex in ice-cold Hanks’ balanced salt solution not containing Ca/Mg (FUJIFILM Wako Chemicals). The cerebrum was suspended in Hanks’ balanced salt solution containing 0.25% trypsin, 0.1 μl/ml deoxyribonuclease (Nippon Gene), 0.8 mM MgSO₄ (Kanto Chemical Co), and 1.85 mM CaCl₂ (Kanto Chemical Co) at 37 °C for 15 min. The obtained cell suspension was passed through a 100 μm cell strainer (Falcon) and centrifuged with a culture medium. The cell pellet was resuspended in the culture medium and seeded on cell culture plates. The culture medium was replaced at 4 days in vitro. The enrichment of glial cells was confirmed by immunocytochemistry using anti-glial fibrillary acidic protein and anti-Iba1 antibodies (Fig. S5).

Neopentyl glycol diacrylate, 1,9-bisacryloyloxynonane, tetrahexylammonium bromide, and diethylenetriamine (DETA) were purchased from Tokyo Chemical Industry (Tokyo, Japan). Et₂O, MgSO₄, hexane, 5% NaOCl aq., and tetrahydrofuran (THF) were purchased from Kanto Chemical (Tokyo, Japan). All the reagents were used as received.