Midbrain organoid astrocyte seeding
gustavo.parfitt Parfitt
Abstract
Midbrain organoid astrocyte seeding protocol
Steps
Refer to the publish protocols to obtain the cell materials required.
https://www.protocols.io/view/midbrain-organoid-differentiation-in-spinner-flask-rm7vzbnr4vx1/v1
https://www.protocols.io/view/astrocyte-extraction-from-brain-organoids-261ge364wl47/v2
From a 90% confluent plate of astrocytes.
Wash with PBS.
Add 1 mL of TrypLE™ Select Enzyme (1X), no phenol red Thermo Fisher Catalog #12563011.
Transfer the cells to 15 ml falcon tubes and add 5 ml of media.
300rcf,25°C
Resuspend the cell pellet in 1ml of
Count the cells using
Equipment
| Value | Label |
|---|---|
| Countess II | NAME |
| Life Technologies | BRAND |
| AMQAX1000 | SKU |
| https://www.thermofisher.com/us/en/home.html | LINK |
Individually add day 40 midbrain organoids to a Corning 96 well V bottom shape plate.
Add 50K astrocyte to the organoids.
Add 100 ul of organoid media.
After a week transfer the organoids to a 96 well plate coated with
