IntestiCult™ Organoid Growth Medium (Human)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Defrost the Matrigel in advance on ice or in fridge	Change medium every 4 days Organoid passage every 2–3 days

Upstream tips
Defrost the Matrigel in advance on ice or in fridge
Protocol tips
Change medium every 4 days Organoid passage every 2–3 days

Publication protocol

GC Organoid Culture: The resuspension was mixed with Matrigel at the ratio of 2: 1. The mixed liquid was added into prewarmed 6-well suspension culture plates with each drop of 40 μL and these plates were then put into an incubator chamber at 37°C for 10 minutes. Upon gelation completed, 1.5 mL IntestiCult Organoid Growth Medium (Stemcell, 06010) was added to each well. Then the plates were transferred to humidified 37°C/5% CO2 incubators. Medium was changed every 4 days and organoids were passaged every 2–3 days. When passaging, cystic organoids were resuspended in 2 mL cold AdDF+++ and mechanically sheared through flamed glass Pasteur pipettes. Dense organoids were resuspended in 2 mL TrypLE Express (Invitrogen, 12,605,036) and incubated for 1–5 min at room temperature. Later, the organoids were mechanically sheared through flamed glass pasteur pipettes. Following the addition of 10 mL AdDF+++ and centrifugation at 300 rcf, dissociated organoids were resuspended in cold Matrigel and seeded as above at ratio1:2 to 1:3. Single cell suspension was initially seeded at a higher density and reseeded at a lower density after one week. Mycoplasma tests were done with the MycoAlert mycoplasma detection kit (Lonza, LT07-318) and all were negative.



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Manufacturer protocol

Download the product protocol from STEMCELL technologies for IntestiCult™ Organoid Growth Medium (Human) below.

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