Preparation of ColoUp medium

The ColoUp medium for cultivation of HCEC-1CT cells contains the herein described components and is prepared as described in the following:

take one bottle (500 ml) of DMEM (Gibco, Cat # 61965-026) and discard 100 mladd 100 ml of M199 (Gibco, Cat# 31150022) and mix properlydiscard 11.6 ml DMEM/M199 mixtureadd 10 ml Cosmic Calf Serum (e.g. Hyclone, Cat# SH30087)add 500 µl of hEGF stock (20 µg/ml, prepared in cell culture grade water, Sigma-Aldrich, Cat# E9644)add 500 µl of Insulin (10 mg/ml, ready-to-use, Sigma-Aldrich, Cat# I9278)add 100 µl of Apo-Transferrin stock (10 mg/ml, prepared in M199, Sigma-Aldrich, Cat# T2036)add 25 µl of Sodium-Selenite stock (100 µM, prepared in cell culture grade water, Sigma-Aldrich, Cat# S5261)add 500 µl Hydrocortisone stock (1 mg/ml, prepared in cell culture grade water, Sigma-Aldrich, Cat# H0396)mix properly and store at 4°C for up to 1 monthtemper the medium to room temperature (not 37°C) before use

Storage conditions

Store the basal medium at 4°C for up to 6 months and the supplements -20°C for up to 6 months.

Store the ready-to-use medium in aliquots at 4°C for up to 1 month.. Temper the medium to room temperature (not 37°C) before use. Avoid repeated heating to 37°C.

Protocols