Mesenchymal Stem Cell Differentiation-Media  22.04.2003

Osteoblast-Medium[1]

50µg/ml  Ascorbic Acid

3mM      b-Glycerophosphate

10nM     Dexamethason

9µg/ml   Oxytetracyclin[2] (fakultative)

in a-MEM with 15% FBS

Dexamethason Stock-Solution (c=10µM)

10µl     4mg/ml Dexamethason-Lösung       (Dexa 4mg inject Jenapharm)

9990µl   a-MEM

Preparation

25mg     Ascorbinsäure

445.5mg  b-Glycerophosphat

500µl    Dexamethason-Stammlösung (c = 10µM = 4mg/l)

45µl     Oxytetracyklin-Stammlösung 10mg/ml in DMSO

ad 500ml a-MEM with 15% FBS

Note

3mM b-Glycerophosphate is a relatively low concentration. More common is 10mM[3].

 

Chondrocyte-Medium[4]

a-MEM w/o FBS

100nM    Dexamethason

10ng/ml  TNF-b3

50µg/ml  Ascorbic acid

100µg/mlNa-Pyruvate

1x       SITE Liquid Media Supplement     Sigma S-4920

1.5mg/mlbovine serum albumin

Adipocyte-Medium[5]

a-MEM, 15%FBS

1µM      Dexamethason

100µg/ml1-methyl-3-isobutylxanthine




[1] M. Ahmad, et al., An in vitro model for mineralization of human osteoblast-like cells on implant materials. Biomaterials, 1999. 20: 211-20.

[2] R. Todescan, et al., Tetracycline fluorescence: a new method to quantify bone produced in vitro. 5th World Biomaterials Congress, 1996: 721.

K. Matsuzaka, et al., The effect of poly-L-lactic acid with parallel surface micro groove on osteoblast-like cells in vitro. Biomaterials, 1999. 20: 1293-301.

[3] M.J. Coelho and M.H. Fernandes, Human bone cell cultures in biocompatibility testing. Part II: effect of ascorbic acid, b-glycerophosphate and dexamethasone on osteoblastic differentiation. Biomaterials, 2000. 21: 1095-102.

[4] B. Johnstone, et al., In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res, 1998. 238: 265-72., A.M. Mackay, et al., Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow. Tissue Eng, 1998. 4: 415-28.

[5] M.E. Nuttall, et al., Human trabecular bone cells are able to express both osteoblastic and adipocytic phenotype: implications for osteopenic disorders. J Bone Miner Res, 1998. 13: 371-82., P.A. Conget and J.J. Minguell, Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells. J Cell Physiol, 1999. 181: 67-73.