Forma series 2 3111 water jacketed co2 incubator

MC3T3-E1 osteoblasts (MC3T3-E1 subclone 4, ATCC CRL2593, USA) were cultured at 37 °C in a 5% CO₂ incubator (Forma Series II 3111 Water Jacketed CO₂ Incubator, Thermo Fisher Scientific Inc., USA) using an alpha minimum essential medium (α-MEM; Gibco-BRL, Grand Island, USA) containing 10% fetal bovine serum (FBS) and 100 U/mL penicillin.

We commercially purchased MC3T3-E1 cell (MC3T3-E1 Subclone 4, ATCC CRL2593, Rockville, MD, USA) from American Type Culture Collection (Manassas, VA, USA). Compared with primary cell cultures, the clonal mouse pre-osteoblastic MC3T3-E1 cell line exhibits high levels of cellular differentiation and clear reproducibility.13 (link) MC3T3-E1 cells are similar to osteoblasts.14 (link) In particular, the MC3T3-E1 Subclone 4 cells represent a good model for studying in vitro osteoblast differentiation.13 (link) In addition, because of behavior similar to primary calvarial osteoblasts, they were considered appropriate for studying the impact of the implant surface on osteoblasts.13 (link) The osteoblastic cell line MC3T3-E1 was dispensed into α-Minimum Essential Medium (α-MEM, Dulbecco’s Modified Eagle Medium, Gibco-BRL, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS) and 100 U/mL penicillin at a concentration of 1 x 10⁶ cells/mL, and was cultured in a carbon dioxide incubator set at 5% CO₂ and 37°C (FormaSeries II 3111 Water Jacketed CO₂ Incubator, Thermo Scientific, Waltham, MA, USA). The cell culture was performed in cell culture dishes (9 x 20 mm). The culture medium was changed every 3 days, and the MC3T3-E1 cells were subcultured 4 times.