Directed differentiation of human embryonic stem cells into the pancreatic endocrine lineage.
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.
Phillips
Blaine W
BW
ES Cell International Pte, Ltd., Singapore 138667.
Hentze
Hannes
H
Rust
William L
WL
Chen
Qi-Ping
QP
Chipperfield
Hiram
H
Tan
Ee-Kim
EK
Abraham
Suman
S
Sadasivam
Akila
A
Soong
Poh Loong
PL
Wang
Siew Tein
ST
Lim
Ricky
R
Sun
William
W
Colman
Alan
A
Dunn
N Ray
NR
D000818:Animals; D002096:C-Peptide / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D017931:DNA Primers; D053595:Embryonic Stem Cells / Q000166:cytology*; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D017403:In Situ Hybridization; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000502:physiology; D051379:Mice; D016133:Polymerase Chain Reaction; D015534:Trans-Activators / Q000235:genetics