The peculiar biology of mouse mesenchymal stromal cells—oxygen is the key

Because of the ability to manipulate their genome, mice are the experimental tool of choice for many areas of scientific investigation. Moreover, established experimental mouse models of human disease are widely available and offer a valuable resource to obtain proof-of-concept for many cell-based therapies. Nevertheless, efforts to establish reliable methods to isolate mesenchymal stromal cells (MSCs) from mouse bone marrow have been elusive. Indeed, a variety of physical and genetic approaches have been described to fractionate MSCs from other cell lineages in bone marrow, but few have achieved high yields or purity while maintaining the genomic integrity of the cells. We provide a historic overview of published procedures dedicated to the isolation of mouse MSCs from bone marrow and compact bone. We also review current findings indicating that growth-restrictive conditions imposed by atmospheric oxygen promotes immortalization of mouse MSCs and how expansion in a low-oxygen environment enhances cell yields and maintains genomic stability. Finally, we provide basic recommendations for isolating primary mouse MSCs and discuss potential pitfalls associated with these isolation methods.