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Isolation of chondrocytes

MN Marcel Niemann
MO Melanie Ort
LL Luis Lauterbach
MS Mathias Streitz
AW Andreas Wilhelm
GG Gerald Grütz
FF Florian N. Fleckenstein
FG Frank Graef
AB Antje Blankenstein
SR Simon Reinke
US Ulrich Stöckle
CP Carsten Perka
GD Georg N. Duda
SG Sven Geißler
TW Tobias Winkler
TM Tazio Maleitzke
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Cartilage was obtained from patients undergoing total knee replacement surgery for knee OA (n=3). Cartilage samples were cut into small pieces, added to the pre-warmed basal medium (low-glucose Dulbecco’s modified Eagle medium [LG-DMEM, Thermo Fisher Scientific Inc., Waltham, MA, USA], 1% penicillin-streptomycin [PenStrep, 10.000 U/ml, Thermo Fisher Scientific Inc.], and 1% GlutaMAX [GlutaMAX™, Thermo Fisher Scientific Inc.]) supplemented with 1mg/ml collagenase type II (Thermo Fisher Scientific Inc.), and incubated at 37 °C for 12–24 h at 5% CO2. Samples were spun down at 300 g for 5 min, supernatants were discarded, and fresh expansion medium (basal medium 10% fetal bovine serum [FBS, Thermo Fisher Scientific Inc.]) was added. The medium was changed every 4 days and cells were grown for 2 to 3 weeks. For all in vitro experiments, cells were used in passages two to three.

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