Isolation and culture of rodent bone marrow-derived multipotent mesenchymal stromal cells

Initial attempts to cultivate mesenchymal stem cells (MSCs) were more successful from human than murine tissues. Methods for the in vitro expansion of murine MSCs were described more recently, but are now well established. Despite limitations such as a poor equivalence to be expected between cultured stem cells and their in vivo counterparts, MSC culture allows the expansion of a cell population capable of providing important information on the biology of stem cells and their therapeutic application. Murine MSCs may be obtained from the bone marrow and virtually from any other organ or tissue. This chapter describes the most widely used method, which involves the preparation of single-cell suspension followed by incubation for 1-3 days and removal of nonadherent cells. The adherent fraction is then expanded by continuous culture and may be maintained for prolonged periods of time.