By Dr. Jennie P. Mather (Eds.)

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2007), on both feeder coculture and feeder-free culture. Morphologically, the hESC retained the undiVerentiated phenotype and pluripotency was confirmed by the positive detection of cell surface markers and intracellular transcription factors. Furthermore, the hESC cultures maintained a normal karyotype (46 X,X) and formed teratomas when injected into severe combined immunodeficient (SCID) mouse. II. Methods and Materials A. Culture of hESC hESC lines are available from a variety of diVerent sources.

Scale bar = 30 mm. E1, was shown to support the undiVerentiated growth of hESC in both feeder and feeder-free conditions. This cell line was previously derived by Lian et al. (2007) and was shown to have similar morphology, phenotype, and function compared to adult MSC. The cells are also 27 2. Autogeneic Feeders for the Culture of UndiVerentiated hESC A 1 2 3 6 7 8 13 14 15 19 20 1 2 3 6 7 8 13 14 15 19 20 9 21 4 5 10 11 12 16 17 18 X Y 22 B 9 21 4 5 10 11 12 16 17 18 X Y 22 Fig. E1 (B). Normal karyotype (46 X,X) was observed for human embryonic stem cells (hESC) from both conditions after six passages.

Membrane-Based Embryonic Stem Cell Culture 31 (Fuller and Simons, 1986; Mostov and Deitcher, 1986). In addition to benefiting the diVerentiation of epithelial cells, epithelial monolayers grown on porous membranes form an eVectively separated dual chamber system for the study of molecular transport across the monolayer. The ability to sample from both separate chambers allows for measurement of the transport of nutrients and drug compounds from one chamber through the cell monolayer to the other chamber in a controlled, in vitro fashion.

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