Cortical bone-derived stem cell culture: cortical bone-derived stem cells were isolated from the tibias and femurs of C57BL/6 mice, as previously described [13 (link),21 (link)]. Briefly, tibias and femurs were flushed to remove all the bone marrow and then digested in collagenase at 37 °C. The digested cells were washed and plated in CBSC media until colonies of CBSCs appeared. The cells were characterized for CBSC markers and expanded for experiments.
Mesenchymal stem cell culture: the femur and tibia of C57BL/6J mice ranging from 2 to 6 months of age were isolated. The epiphysis of each were removed and each bone cavity was flushed with pre-warmed, and the complete culture medium for bone marrow extracted. Extracted marrow medium was passed through a 70 μM filter and remaining cell suspension was cultured in DMEM (Corning) containing 20% fetal bovine serum (Neuromics, Edina, Min, USA) and 1% penicillin streptomycin/L-glutamine (Gibco Life Technologies, Waltham, MA, USA). MSCs were isolated from other bone marrow cells via two passages of plastic adherence. Remaining cells were expanded and cryopreserved as previously described [40 (link)].
Cardiac-derived stem cells culture: cardiac-derived stem cells RNA and protein were generously gifted to us from Dr. Mohsin Khan’s laboratory.
Mesenchymal stem cell culture: the femur and tibia of C57BL/6J mice ranging from 2 to 6 months of age were isolated. The epiphysis of each were removed and each bone cavity was flushed with pre-warmed, and the complete culture medium for bone marrow extracted. Extracted marrow medium was passed through a 70 μM filter and remaining cell suspension was cultured in DMEM (Corning) containing 20% fetal bovine serum (Neuromics, Edina, Min, USA) and 1% penicillin streptomycin/L-glutamine (Gibco Life Technologies, Waltham, MA, USA). MSCs were isolated from other bone marrow cells via two passages of plastic adherence. Remaining cells were expanded and cryopreserved as previously described [40 (link)].
Cardiac-derived stem cells culture: cardiac-derived stem cells RNA and protein were generously gifted to us from Dr. Mohsin Khan’s laboratory.
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