Isolation and Culturing of Cells from Mouse Embryo Dorsolateral Telencephalons

Abstract

Source: Landeira, B. S., et al. Live Imaging of Primary Cerebral Cortex Cells Using a 2D Culture System. J. Vis. Exp. (2017)

This video demonstrates a protocol for isolating and culturing primary cerebral cortex cells from the dorsolateral telencephalon of mouse embryos.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Dorsolateral Telencephalon Microdissection

1. Prepare the dissection medium (100 mL): 98.5 mL Hank's Balanced Salt Solution (HBSS), 0.5 mL 5 M 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) pH 7.4, 1 mL penicillin/streptomycin (10,000 units/mL and 10,000 µg/mL).
2. Filter-sterilize the dissection medium.
3. Remove embryonic day 14 (E14) embryos from a pregnant mouse C57/Bl6 (Mus musculus) under anesthesia with iso....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Hank's Balanced Salt Solution (HBSS)	Invitrogen Life Technologies	14175129
HEPES	Sigma-Aldrich	H3375-25G
Penicillin/streptomycin	Gibco	15140122
Dulbecco Modified Eagle's Medium (DMEM)	Gibco	12400-024
Fetal Calf Serum (FCS)	Gibco	10437028
Glucose	Gibco	A2494001
B27	Gibco	17504044
Trypsin-EDTA (0.05%)	Gibco	25300054
Cell observer microscope	Zeiss