Cell adhesion, morphology of ES cells on the E-cad-Fc fusion protein-immobilized surface.
(A) ES cells (EB3) adhered to E-cad-Fc-coated dishes with equivalent efficiency as to 0.1% gelatin-coated dishes after 3 hours of incubation.
(B) ES cells (EB3) were cultured on E-cad-Fc-coated or fibronectin-coated dishes without serum.
EGTA (5 mM) was added to the culture medium at 3 hours after seeding (open bar).
Detached cells were removed and remaining cells were counted using alamar Blue reagent.
*:P<0.05, §:P<0.001 vs. no treated condition (closed bar).
(C and D) Morphological observation of ES cells (EB3) on the two different matrices.
ES cells were cultured on polystyrene surfaces coated with 0.1% (wt/vol) gelatin (C), or 10 µg/ml E-cad-Fc (D) in the presence of LIF for 2 days.
High magnification images are shown in (C′) and (D′).
(E) ES cells (EB3 and R1 cells) were cultured on the plates coated with gelatin or E-cad-Fc and differentiation was induced by the withdrawal of LIF.
Morphological characteristics were observed as phase contrast images.
Bar indicates 100 µm.
The data indicate means±SD of 3 separate experiments.