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: