Dimethyloxaloylglycine increases bone repair capacity of adipose-derived stem cells in the treatment of osteonecrosis of the femoral head

Zhu, Zhen-Hong; Song, Wen-Qi; Zhang, Chang-Qing; Yin, Ji-Min

Isolation and culture of ASCs. Primary ASCs were harvested from the adipose tissue of New Zealand rabbits. Briefly, the animals were anesthetized with pentobarbital sodium (3 mg/100 g; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany). Adipose tissues were harvested and digested with 0.1% collagenase I (Sigma-Aldrich) for 1 h. The complex was filtered with a 100-µm nylon mesh (Shanghai Bolting Cloth Manufacturing Co., Ltd., Shanghai, China) and centrifuged at room temperature for 30 min at 363 x g The cells were then resuspended with Dulbecco's modified Eagle's medium (Gibco; Thermo Fisher Scientific, Inc., Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Invitrogen; Thermo Fisher Scientific, Inc., Carlsbad, CA, USA), and were plated on culture flask (Corning Life Sciences, Tewksbury, MA, USA). The cells were cultured at 37˚C in a humidified 5% CO 2 incubator. The culture medium was replaced every 3 days, and non-adherent cells were removed. The cells were passaged approximately at a 1:3 split at subconfluence. The cells of four to six passages were used for the following experiments.