Chondrogenic
Media II
2010, Dominik Haudenschild, UC Davis
Chondrogenic Base Media II
permissive of chondrogenesis
á DMEM/F12, 3.1g/L Glucose
o 4mM L-Glutamine
o 100µg/ml Na-Pyruvate
á 15mM HEPES pH 7.5
á 40µg/ml L- Proline
á 50µg/ml L-ascorbic acid-2-phosphate
á 100nM Dexamethasone
á 1% ITS+
á
{+10mM
CaCl2 and 50µM MnCl2 for Alginate Culture}
Chondrogenic Media IIa
promotes chondrogenesis
á Base chondrogenic Media
o + 10ng/ml TGF-§1
Chondrogenic Media IIb
strongly promotes chondrogenesis
á Base Chondrogenic Media
o + 10ng/ml TGF-§1
o + 100ng/ml BMP-7
500ml bottle (HyClone SH30023.01, contains some glu-pyr)
add 10ml of
200mM glutamine (Invitrogen 25030-081)
add 5ml of
100mM pyruvate (GIBCO 11360, VMCS GB132)
add 1ml of 20mg/ml proline (Sigma P-5607, in H20, FS)
add 1ml of 25mg/ml ascorbic acid (Sigma A8960, in H20, FS)
add 1ml of 1:150 dilution of 4mg/ml dexamethasone
add 5.5ml of ITS+ (Clontech 354352)
add 5ml 100x Cation solution, final is 10mM Ca, 50µM Mn
add 2.5ml 10mM MnCl2 (final
= 50µM Manganese)
NOTE:
DME/F12 chosen over DME because it contains Zn
100x Cation Solution
Contains:
8.89M
CaCl2 and 5mM MnCl2
Link
to Chondrogenic Supplement
that contains all of above: