Fig. 4.

EBs from the 4 hESC lines; SA002, AS034, SA121 and SA167 cultured in agarose for 8 weeks developed heterogeneous differentiation patterns. Histological analysis with Alcian Blue-van Gieson staining showed differentiation heterogeneity both between EBs from the 4 hESC lines and between different EBs from the same hESC line. EBs from the AS034, SA121 and SA167 in three individual replicates developed highly differentiated areas with cell types from the three germ layers (thin black arrow for neuronal rosettes like structures representing ectoderm, white arrow for connective tissue representing mesoderm and thick black arrow for epithelia representing endoderm). The EBs from the SA002 cell line did not develop highly differentiated structures but produced extracellular matrix. Bar 100 μm