For in-vitro study, human mesenchymal stem cells (hMSCs, RoosterBio Inc., MD, USA) were cultured using MSC expansion media (R&D Systems, MN, USA) (growth media; GM) supplemented with Penicillin (100 IU/ml) - Streptomycin (100 μg/ml) (cat. no. 30–002-CI, Corning Life Sciences, Lowell, MA) until usage. As an osteogenic media (OM), human osteogenic differentiation media (Cell Applications, USA) added with Penicillin-Streptomycin was used.
For in-vivo study, as implantation of hMSCs was not possible into immunocompetent rats, allogenic rBMSCs were extracted from inbred 4-week-old male Fischer white rats (F344/DuCrl, Charles River Laboratories, Wilmington, MA) [25 ]. All animals were cared in the animal facility (Millennium Science Complex, PSU) and euthanasia procedures were according to American Association for Laboratory Animal Science (AALAS) and The Institutional Animal Care and Use Committee (IACUC protocol #46591). Isolated rBMSCs were plated on 6-well cell culture plates in Minimal Eagle’s Medium, Alpha modification (αMEM; cat. no. 10–022-CV, Corning Cellgro®, Manassas, VA) supplemented with 10% fetal bovine serum (FBS) (cat. no. 35–010-cv, Thermo Fisher Scientific, Waltham, WA), PS and 2.5 μg/ml Fungizone (cat. no. 15–290-026, Thermo Fisher Scientific) and incubated at 37 °C in a humidified 5% CO2 incubator.
For in-vivo study, as implantation of hMSCs was not possible into immunocompetent rats, allogenic rBMSCs were extracted from inbred 4-week-old male Fischer white rats (F344/DuCrl, Charles River Laboratories, Wilmington, MA) [25 ]. All animals were cared in the animal facility (Millennium Science Complex, PSU) and euthanasia procedures were according to American Association for Laboratory Animal Science (AALAS) and The Institutional Animal Care and Use Committee (IACUC protocol #46591). Isolated rBMSCs were plated on 6-well cell culture plates in Minimal Eagle’s Medium, Alpha modification (αMEM; cat. no. 10–022-CV, Corning Cellgro®, Manassas, VA) supplemented with 10% fetal bovine serum (FBS) (cat. no. 35–010-cv, Thermo Fisher Scientific, Waltham, WA), PS and 2.5 μg/ml Fungizone (cat. no. 15–290-026, Thermo Fisher Scientific) and incubated at 37 °C in a humidified 5% CO2 incubator.