OSCM — Organotypic Slice Culture Medium (1 L)

Defined serum-free medium for ex vivo organotypic human brain slice culture. Hover any reagent for vendor, catalog #, storage, and (eventually) location.

pH7.35–7.45

Osmolarity300–320 mOsm

Shelf life2 weeks at 4°C

Add components in the order listed. The 300 mL initial water is a dissolving solvent; the final ~600 mL should be added in smaller portions, titrating to reach osmolarity 300–320 mOsm.

Recipe

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#	Component	Final conc.	Volume (1 L)	Notes
1	BME	dilute to 15%	150 mL
2	DMEM/F12	dilute to 15%	150 mL
3	H₂O (initial)	—	300 mL	Initial dissolving solvent
4	CaCl₂	0.91 mM	1 mL of 1000× aliquot
5	KCl	1.96 mM	26.15 mL	Ready-made, not aliquoted
6	MgSO₄	0.81 mM	1 mL of 1000× aliquot
7	NaCl	81.90 mM	16.40 mL
8	NaH₂PO₄	0.84 mM	1 mL of 1000× aliquot
9	NaHCO₃	14.00 mM	4.00 mL
10	ZnSO₄	0.5 μM	0.05 mL (50 μL)
11	CuSO₄	0.01 μM	1 mL of 1000× aliquot
12	D-Glucose	0.63 mM	1 mL of 1000× aliquot
13	Sodium pyruvate	0.14 mM	1.40 mL
14	Ascorbic acid	0.58 mM	1 mL of 1000× aliquot
15	Oxalo-acetic acid	1.00 mM	1 mL of 1000× aliquot
16	Sodium lactate	4.00 mM	4.00 mL
17	Citric acid	0.57 mM	1 mL of 1000× aliquot
18	Acetic acid	0.57 mM	0.03 mL (30 μL)	Glacial, from benchtop
19	BSA	0.265%	26.50 mL
20	17-β-Estradiol	0.001 μM	1 mL of 1000× aliquot
21	α-Tocopherol	2.28 μM	1 mL of 1000× aliquot
22	α-Tocopherol acetate	2.10 μM	1 mL of 1000× aliquot
23	Corticosterone	0.06 μM	1 mL of 1000× aliquot
24	Creatine	5.00 mM	655 mg (powder)	Weigh direct; or 50 mL of 0.1 M stock
25	Sodium β-hydroxybutyrate	1.00 mM	1 mL of 1000× aliquot
26	Mannitol	5.00 mM	10.00 mL
27	Phosphate creatine	0.2 mM	1 mL of 1000× aliquot
28	N-2 supplement	0.5% (1×)	5.00 mL	Ready-made, not aliquoted
29	ATP	0.01 mM	1 mL of 1000× aliquot
30	Insulin	0.41 μM	0.60 mL	Ready-made, not aliquoted
31	Tri-iodo-L-thyronine (T3)	0.0003 μM	100 μL of 10,000× aliquot
32	Superoxide dismutase	0.08 μM	1 mL of 1000× aliquot
33	Glycerol	0.05 mM	0.50 mL
34	GlutaMax	0.5 mM	2.50 mL	Ready-made, not aliquoted
35	MnCl₂	1.00 μM	100 μL of 10,000× aliquot
36	γ-Amino-butyric acid (GABA)	1.00 μM	100 μL of 10,000× aliquot
37	Glutathione	3.00 μM	100 μL of 10,000× aliquot
38	Taurine	8 μM	80 μL of 10,000× aliquot
39	HEPES	5 mM	1,191 mg (powder)	Weigh direct; or 5 mL of 1 M stock
40	H₂O (final, titrated)	—	~600 mL	Add in smaller portions; titrate to osm 300–320 mOsm
Final volume:			1,000 mL (1 L)

Notes

Lab aliquot convention: most −20°C reagents that aren't commercially ready-made are pre-aliquoted at 1000× (add 1 mL per 1 L recipe). A handful of small-volume reagents are pre-aliquoted at 10,000× (add 100 μL per 1 L recipe) — flagged in the Volume column.
Ready-made exceptions (not aliquoted): BME, DMEM/F12, H₂O, KCl, NaCl, NaHCO₃, ZnSO₄, Sodium pyruvate, Sodium lactate, BSA, Mannitol, N-2 supplement, Insulin, Glycerol, GlutaMax, Acetic acid.
Powder reagents (Creatine, HEPES): weigh directly for 1 L recipe rather than making a stock.
Removed from original recipe: Heparin (Sigma H3149) — struck out; not included.
Scale: published Nature Protocols recipe is for 100 mL; this page shows the 1 L version.
Checklist: checkbox state is saved in your browser only (localStorage), per-device. Won't sync between people. Refresh-safe. Hit "Reset checklist" when you start a fresh prep.

Source

Adapted from Table 2 of: Park TI, Smyth LCD, Aalderink M, Woolf ZR, et al. Routine culture and study of adult human brain cells from neurosurgical specimens. Nature Protocols 17, 190–221 (2022). doi:10.1038/s41596-021-00637-8 · PMID 35022619