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Directed Differentiation of Human Embryonic Stem Cells into the Pancreatic Endocrine Lineage

2007; Mary Ann Liebert, Inc.; Volume: 16; Issue: 4 Linguagem: Inglês

10.1089/scd.2007.0029

1557-8534

Blaine Phillips, Hannes Hentze, William L. Rust, Qi-Ping Chen, Hiram Chipperfield, Ee-Kim Tan, Suman Abraham, Akila Sadasivam, Poh Loong Soong, Siew Tein Wang, Ricky Yee Meng Lim, William Sun, Alan Colman, N. Ray Dunn,

Congenital heart defects research

Human embryonic stem (hES) cells represent a potentially unlimited source of transplantable beta-cells for the treatment of diabetes. Here we describe a differentiation strategy that reproducibly directs HES3, an National Institutes of Health (NIH)-registered hES cell line, into cells of the pancreatic endocrine lineage. HES3 cells are removed from their feeder layer and cultured as embryoid bodies in a three-dimensional matrix in the presence of Activin A and Bmp4 to induce definitive endoderm. Next, growth factors known to promote the proliferation and differentiation of pancreatic ductal epithelial cells to glucose-sensing, insulin-secreting beta-cells are added. Pdx1 expression, which identifies pancreatic progenitors, is detected as early as day 12 of differentiation. By day 34, Pdx1+ cells comprise between 5% and 20% of the total cell population and Insulin gene expression is up-regulated, with release of C-peptide into the culture medium. Unlike another recent report of the induction of insulin+ cells in differentiated hES cell populations, we are unable to detect the expression of other pancreatic hormones in insulin+ cells. When transplanted into severe combined immunodeficiency (SCID) mice, differentiated cell populations retain their endocrine identity and synthesize insulin.