Magnetic cell sorting isolation of therapeutically effective BALB/c mouse bone marrow hematopoietic stem cell population

Abstract

Purified bone marrow cells are being successfully used to restore the injured tissue and may serve as an alternative to organ transplantation. From the therapeutic point of view, it is of importance to obtain individual immunophenotypically characterized hematopoietic cell populations which, after their predisposed differentiation performed in vitro, might be applied selectively. In this paper, we demonstrate, BALB/c mice using as a model, how a therapeutically effective bone marrow hematopoietic stem cell population can be easily isolated by magnetic cell sorting techniques. Hematopoietic progenitor stem cells were isolated from bone marrow mononuclear cell suspension by negative selection (differentiated cells were removed using nanoparticles labeled with antibodies against Lin+). Hematopoietic stem cells of interest were isolated from Lin- (negative fraction) using BD IMagTM anti-mouse CD117 particles (positive selection). Flow cytometrical analysis of these cells revealed that 42.59 ± 5.74% of cells expressed CD38+CD117+, 3.59 ± 0.84% expressed CD34–CD117+, and 60.94 ± 1.43% expressed Sca1+CD117+. The cell population was therapeutically highly effective when examined in a contact hypersensitivity experiment. We believe that the methodological approach described in this paper could be easily extended in regard to therapeutically important stem cells of other origins. Keywords: BALB/c mouse, bone marrow, magnetic cell sorting, flow cytometry