Directed differentiation of human embryonic stem cells into the pancreatic endocrine lineage.

Abstract:

: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.

journal_name

Stem Cells Dev

authors

Phillips BW,Hentze H,Rust WL,Chen QP,Chipperfield H,Tan EK,Abraham S,Sadasivam A,Soong PL,Wang ST,Lim R,Sun W,Colman A,Dunn NR

doi

10.1089/scd.2007.0029

subject

Has Abstract

pub_date

2007-08-01 00:00:00

pages

561-78

issue

4

eissn

1547-3287

issn

1557-8534

journal_volume

16

pub_type

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