Peter van Mourik

76 Chapter 4 MATERIALS AND EQUIPMENT: Culture Medium Abbreviation Description Application DMEM +/+ DMEM +/+ Dulbecco’s Modified Eagles Medium with additional 1 % (v/v) pen- strep (10,000 U/mL) and 10 % (v/v) FBS Preparation of conditioned media Ad-DF+++ Ad-DF+++ Advanced Dulbecco’s Modified Eagles Medium with Nutrient Mixture F-12 Hams (Ad-DF 500 mL + 1% (v/v) glutamax, 1% (v/v) hepes buffer and 1% (v/v) pen-strep (see table 3 for exact concentrations) Preparation of conditioned media, handling of colon organoid cultures. Human colon organoid medium -/- 2x CM -/- Colon organoid culture medium with out WCM & R-spo3 (see stock preparation of human colon organoid medium components) Preparation of 50 mL stock 2x human colon organoid medium saved at -20°C Human colon organoid medium +/+ CM +/+ Colon organoid culture medium with WCM & R-spo3 (see stock preparation of human colon organoid medium components) Human colon organoid medium used for regular culture and FIS Noggin conditioned medium NCM Self-produced Noggin conditioned medium 10% of human colon medium Wnt-3A conditioned medium WCM Self-produced Wnt-3a conditioned medium 50% of human colon medium +/+