Human MSC of non-osteoporotic donors were obtained from bone marrow of femoral heads according to the described protocol [53] (link) after total hip arthroplasty due to osteoarthritis and/or hip dysplasia. MSC of patients suffering from osteoporosis were isolated from femoral heads after low-energy fracture of the femoral neck. Additional criteria for confirming primary osteoporosis in these donors were vertebrae fractures and advanced age.
Cell culture medium, fetal calf serum (FCS), trypsin-EDTA and antibiotics were obtained from PAA Laboratories GmbH, Linz, Austria. Human MSC were selected by surface adherence and expanded in DMEM/Ham's F-12 (1∶1) medium supplemented with 10% heat-inactivated FCS, 1 U/ml penicillin, 100 µg/ml streptomycin and 50 µg/ml L-ascorbic acid 2-phosphate (Sigma Aldrich GmbH, Schnelldorf, Germany).
For long term cultivation, cells were expanded at 70–90% confluence by trypsinization with 1× trypsin-EDTA and reseeding in a ratio of 1∶3. This procedure was repeated for up to x passages when the hMSC did not become confluent within 3 weeks due to replicative senescence.
Cell culture medium, fetal calf serum (FCS), trypsin-EDTA and antibiotics were obtained from PAA Laboratories GmbH, Linz, Austria. Human MSC were selected by surface adherence and expanded in DMEM/Ham's F-12 (1∶1) medium supplemented with 10% heat-inactivated FCS, 1 U/ml penicillin, 100 µg/ml streptomycin and 50 µg/ml L-ascorbic acid 2-phosphate (Sigma Aldrich GmbH, Schnelldorf, Germany).
For long term cultivation, cells were expanded at 70–90% confluence by trypsinization with 1× trypsin-EDTA and reseeding in a ratio of 1∶3. This procedure was repeated for up to x passages when the hMSC did not become confluent within 3 weeks due to replicative senescence.
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