Immunomagnetic Isolation of Mouse Embryonic Stem Cells from Heterogeneous Cell Population

A simple immunomagnetic cell sorting method is described for the isolation and the purification of mouse embryonic stem (ES) cells from a heterogeneous cell mixture containing both ES cells and mouse embryonic fibroblast cells (MEF). The cells (1.5 × 106 cells per experiment), which labelled with the monoclonal SSEA-1 (stage specific embryonic antigen 1) antibody followed by the microbead coated with anti mouse IgM antibody, were retained in the separation column held in a magnetic field. Over 95% of the cells collected were viable after immunomagnetic cell sorting. Recoveries of 68, 82 and 90% and purity 83, 82 and 73% of the ES cells were obtained using 10, 20 and 50 μg of SSEA-1 antibody per experiment, respectively. Procedure for isolation did not affect the proliferation of the isolated cells on MEF in culture. These results suggest that the immunomagnetic cell sorting using undifferentiated cell marker antibody could be highly effective for isolating the undifferentiated cells such as ES cells.