Establishment and application of cell cultures from mouse bone explants

Abstract

The aim of our study was to establish primary cell cultures from mouse bone explants and to introduce them in our laboratory as routine model system in biocompatibility assessment of new materials for bone implants. The cultures were obtained from ICR mice both sexes. Some of their characteristics were studied such as optimal growth conditions (cell culture media and supplements), morphology, in vitro growth properties (doubling time, plating efficiency, ability to form 3D colonies in soft agar), tumorigenic potential in vivo. The cells were maintained as adherent cultures in a Dulbecco`s modified Eagle`s medium (D-MEM) supplemented with 10% fetal bovine serum and antibiotics penicillin (100 U/mL) and streptomycin (100 mg/mL) at standard concentrations. For routine passages the cells were detached using a mixture of 0.05% trypsin and 0.02% EDTA. Some of the cultures survived for more than 50 passages and are still under cultivation. The cells did not form 3D colonies in semi-solid medium and did not induce tumor growths when implanted s.c. into the back of new born ICR mice (7 x 106 cells/mouse). The cultures were successfully applied in biocompatibility assessment of new materials (obtained from bacterial cellulose) for bone implants.