50µg/ml Ascorbic Acid
3mM b-Glycerophosphate
10nM Dexamethason
9µg/ml Oxytetracyclin[2] (fakultative)
in a-MEM with 15% FBS
10µl 4mg/ml Dexamethason-Lösung (Dexa 4mg inject Jenapharm)
9990µl a-MEM
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
3mM b-Glycerophosphate is a relatively low concentration. More common is 10mM[3].
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
a-MEM, 15%FBS
1µM Dexamethason
100µg/ml1-methyl-3-isobutylxanthine
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[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.