P-EMBL-166 - P-EMBL-166
In this study we analyzed mesenchymal stem cells (MSC) from the bone marrow (BM) that were isolated from the same donors under two different growth conditions (M1 and M2). Bone marrow aspirates were obtained from the iliac crest of four healthy donors aged 25-35 years after approval by the Heidelberg University Ethical Board (approval nos.: 042/2000 and 251/2002). About 10-30 ml bone marrow aspirate were collected in a syringe containing 10,000 IU heparin to prevent coagulation. The mononuclear cell fraction was isolated by Biocoll density gradient centifugation (d=1.077 g/cm3; Biochrom, Germany).
BM-MSC-M2 were cultivated in the commercially available Poietics Human Mesenchymal Stem Cell Medium (M2; PT-3001, Cambrex) following the manufacturer's instructions. About 100,000 cells/cm2 were plated in tissue culture flasks without fibronectin coating. The plastic adherent cell fraction was re-seeded at a density of 5,000 to 6,000 cells/cm2. Cells were expanded for 2-6 passages. Osteogenesis, chondrogenesis and adipogenesis could be induced by the commercially available differentiation media (Cambrex).
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