Embryoid body-mediated differentiation of hiPSCs derived from DPCs in serum-free and feeder-free defined culture conditions and teratoma formation of hiPSCs in the defined culture conditions.

<p>A) Differentiation was performed using embryoid body formation, and the differentiated iPSCs (DP-A-iPS/hESF9 or DP-F-iPS/hESF9T) were fixed and reacted with antibodies. Shown were immunocytochemistry of Nestin, βIII-tubulin, α-smooth muscle actin (α-SMA), and α-fetoprotein (AFP). Binding of these antibodies was visualized with Alexa Fluor 488-conjugated secondary antibodies (green). Oct3/4 was also investigated. Binding of these antibodies was visualized with Alexa Fluor® 594-conjugated secondary antibodies (red). Nucleuses were stained with DAPI. (passage 25). Bar indicates 100 µm. B) Teratomas were generated in SCID mice (CB17/Icr-<i>Prkdc^scid</i>/CrlCrlj) from DP-A-iPS and DP-F-iPS grown under hESF9 or hESF9T-based conditions. Histological analysis with HE staining or Alcian Blue staining demonstrated that teratomas formed from iPS cells cultured in KSR-based (data not shown) or in hESF9T-based conditions contained derivatives of all three germ layers. Left panel shows teratomas from DP-A-iPS-CL1 at passage 22. Right panel shows teratomas from DP-F-iPS-CL14 at passage 6. Scale bars represent 200 µm.