Supplemental Data

Files in this Data Supplement:

• Supplemental Movie 1 - (06-7756SuppMovie1.mpeg; 4.57 MB) RAR antagonist prevents RA-induced aggregate formation by NE-4C neuroectodermal progenitors, as it is shown by long-term time-lapse recordings. The cells were seeded onto poly-L-lysine coated plastic dishes in 10000 cell/cm2 density, in defined medium supplemented with 0.5 % FCS. After the initial attachment, and each day afterwards, the medium was replaced with serum free medium. The videos demonstrate that the NE-4C cells continuously proliferate in control cultures but form dense aggregates in cultures treated with 10 nM RA. 100 nM RAR-antagonist does not alter the behaviour of the NE-4C cells, but effectively prevents aggregate formation induced by 10 nM RA. The antagonist was added to the NE-4C cells 45 minutes prior to RA treatment.
• Supplemental Movie 2 - (06-7756SuppMovie2.mpeg; 6.29 MB) The long-term time-lapse videos demonstrate that the RAR-antagonist prevents astroglia-induced aggregate formation by NE-4C neuroectodermal progenitors. Astrocytes were grown to confluency. Part of the glial sheet was removed 48 hours prior to co-culture establishment. The NE-4C cells were seeded onto confluent monolayers of forebrain astrocytes, in 8000 cell/cm² density, in 0.5 % FCS containing medium. After the initial attachment, and each day afterwards, the medium was replaced with serum free medium. NE-4C cells formed aggregates both in contact and in non-contact areas of the co-cultures. The aggregation of NE-4C cells was effectively prevented by daily application of 100 nM RAR antagonist.
• Supplemental Movie 3A - (06-7756SuppMovie3A.mpeg; 14.4 MB) and Time-lapse recordings show extensive neuronal differentiation by embryonic stem cells in the presence of astroglial cells (CD1-GFP ES). A) Development of a small embryoid body into a large cell cluster on the surface of an astroglial monolayer, demonstrated by a 6 day long recording. See neuronal process outgrowth from the 4th day on.
• Supplemental Movie 3B - (06-7756SuppMovie3B.mpeg; 5.90 MB) B) A part of Movie A that shows details of neuronal outmigration.
• Supplemental Movie 4 - (06-7756SuppMovie4.mpeg; 10.4 MB) The video shows ES cell derived pulsating heart muscle tissue, developed in RAR-antagonist treated ES/astroglia co-culture. The beating tissue is part of a large ES cell cluster.