Human MSCs (hMSCs) were derived from bone marrow samples from 5 healthy premenopausal women (41–46 years old) and 5 women with postmenopausal osteoporosis (51–59 years old). Informed consent was obtained from each participant as a donor of bone marrow. Osteoporosis was diagnosed using the World Health Organization parameters. The hMSCs were purified from the bone marrow samples according to the Percoll density gradient centrifugation method (GE Healthcare Life Sciences). The primary mouse bone marrow cells were collected from the tibia and femur by crushing the bones, then the mouse MSCs (mMSCs) were isolated and purified using EasySep™ Mouse Mesenchymal Stem/Progenitor Cell Enrichment Kit (StemCell). Both hMSCs and mMSCs were cultured in a modified essential medium (a-MEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 U/ml penicillin, and 100 mg/mL streptomycin in an incubator with a humidified atmosphere and 5% CO2 at 37°C. The hMSCs and mMSCs, which were positive (>95%) for CD29, CD44, CD90, and CD105 but were negative (<5%) for hematopoietic markers CD34, CD14, and CD45, were used for further study.
HEK-293T cells were obtained from ATCC and were cultured in DMEM medium supplemented with 10% FBS, 2 mM L-glutamine, 100 μg/mL streptomycin, and 100 U/mL penicillin in an incubator with a humidified atmosphere and 5% CO2 at 37°C.
HEK-293T cells were obtained from ATCC and were cultured in DMEM medium supplemented with 10% FBS, 2 mM L-glutamine, 100 μg/mL streptomycin, and 100 U/mL penicillin in an incubator with a humidified atmosphere and 5% CO2 at 37°C.