Biopharma Insights Here, we describe a method for efficient cryopreservation and thawing of cortical brain tissue blocks to generate highly enriched neuronal cultures. This simple protocol provides flexibility for later generation of neuronal, astrocyte, and neuronal precursor cell cultures.
Total time
~4–5 hours (including cryopreservation); cultures mature over 7–14 days post-thaw
Model organism
Mouse (cortical tissue)
Steps
1
Prepare materials and reagents
Gather and prepare all necessary media, cryoprotectants, and equipment required for tissue cryopreservation and culture.
▶ 01:15
2
Dissect and prepare cortical tissue blocks
Isolate cortical tissue from brain and cut into uniform blocks suitable for cryopreservation.
▶ 02:41
3
Cryopreserve tissue blocks
Equilibrate tissue in cryoprotectant solution and freeze blocks using controlled-rate freezing to preserve cell viability.
▶ 04:20
4
Thaw tissue and establish cultures
Rapidly thaw preserved tissue blocks and plate in culture medium to generate enriched neuronal and glial populations.
▶ 06:05