Preparation of mouse bone marrow primary cultures for sister chromatid exchange and chromosomal aberration studies

Abstract

A procedure for preparing and culturing mouse bone marrow cells for cytogenetic studies is described. Animals are killed by cervical dislocation, the bone marrow is flushed from femora and tibia with Ham's F12 medium into centrifuge tubes. Bone marrow cells are collected by low-speed centrifugation (285 ×g). One million cells are suspended in a 30-ml Falcon flask with complete medium containing 3.45 ml Ham's F12 medium, 1 ml fetal bovine serum, 0.05 ml penicillin-streptomycin, and 0.5 ml whole uterus extract from pregnant mice. For sister chromatid exchange analysis, 20 µM of 5-bromo-2′ -deoxyuridine is also added to the culture medium for 24 to 28 h. The cell cycle is approximately 12 to 14 h, in culture. This culture system can also be used for chromosomal aberration studies.