Figure 1

Human ES cells maintain pluripotency on hE-cad-Fc-coated plates. (A) Purified hE-cad-Fc was separated by SDS-PAGE, and three bands were observed by CBB staining. TOF-MASS analysis revealed that the major 120 kDa band (*) represents full-length hE-cad-Fc, while the 90 kDa band (black arrowhead) was E-cadherin alone and the 30 kDa band (white arrowhead) was the IgG Fc protein alone. (B) Phase contrast micrographs showing the morphology of H9 huES cells cultured on polystyrene surfaces coated with Matrigel or hE-cad-Fc in MEF conditioned medium 7 days after seeding. (C) Histochemistry revealing alkaline phosphatase activity (purple) in H9 cells cultured on Matrigel or on hE-cad-Fc after 68 passages. (D) Immunocytochemistry to detect OCT3/4 (red) in H9 cells cultured on hE-cad-Fc-coated dishes for 13 passages (90 days). Nuclei were counterstained with DAPI. Scale bar indicates 100 μm. (E) The expression of SSEA-4 was analyzed by FACS after 69 passages on hE-cad-Fc. (F) H9 cells were cultured on feeder cells (MEF) or on hE-cad-Fc-coated plates for 13 passages (90 days) (ES), and then in suspension to form embryoid bodies for 21 days (EB). The expression of mRNAs characteristic of the undifferentiated state and specific cell lineages was then identified by RT-PCR.