Muscle

Preparation of MyoUp medium

The MyoUp medium for cultivation of LHCN-M2 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 ml

add 100 ml of M199 (Gibco, Cat# 31150022) and mix properly

discard 90 ml DMEM/M199 mixture

add 75 ml FBS (e.g. PAN Biotech, Cat# P30-3031)

add 10 ml Hepes (1M stock, ready-to-use, Sigma-Aldrich, Cat# H0887)

add 50 µl Zink Sulfate stock (30 mg/ml, prepared in cell culture grade water, Sigma-Aldrich, Cat# Z0251)

add 50 µl Vitamin B12 stock (14 mg/ml, prepared in cell culture grade water, Sigma-Aldrich, Cat# V2876)

add 70 µl Dexamethasone stock (1 mM, prepared in absolute ethanol/DMEM, Sigma-Aldrich, Cat# D4902)

add 25 µl HGF stock (50 µg/ml, prepared in cell culture grade water, Merck Millipore, Cat# GF116)

add 50 µl bFGF stock solution (100 µg/ml, prepared in PBS/Tris/BSA buffer, Peprotech, Cat# 100-18B)

mix properly and store at 4°C for up to 1 month

Storage conditions

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