Immunomagnetic isolation of mouse embryonic stem cells from heterogeneous cell population.

摘要

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 (1.5X 106 cells per experiment) were labelled with the monoclonal SSEA-1 (stage-specific embryonic antigen 1) antibody, and after they had been mixed with microbeads coated with anti-mouse IgM antibody they were retained in a 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% of the ES cells, and purities of 83, 82 and 73%, were obtained using 10, 20 and 50 鎔 SSEA-1 antibody per experiment respectively. Theprocedure for isolation did not affect proliferation of the isolated cells on MEF in culture.