Prospectively isolated NGN3-expressing progenitors from human embryonic stem cells give rise to pancreatic endocrine cells.
Pancreatic endocrine progenitors obtained from human embryonic stem cells (hESCs) represent a promising source to develop cell-based therapies for diabetes. Although endocrine pancreas progenitor cells have been isolated from mouse pancreata on the basis of Ngn3 expression, human endocrine progenitors have not been isolated yet. As substantial differences exist between human and murine pancreas biology, we investigated whether it is possible to isolate pancreatic endocrine progenitors from differentiating hESC cultures by lineage tracing of NGN3. We targeted the 3' end of NGN3 using zinc finger nuclease-mediated homologous recombination to allow selection of NGN3eGFP(+) cells without disrupting the coding sequence of the gene. Isolated NGN3eGFP(+) cells express PDX1, NKX6.1, and chromogranin A and differentiate in vivo toward insulin, glucagon, and somatostatin single hormone-expressing cells but not to ductal or exocrine pancreatic cells or other endodermal, mesodermal, or ectodermal lineages. This confirms that NGN3(+) cells represent pancreatic endocrine progenitors in humans. In addition, this hESC reporter line constitutes a unique tool that may aid in gaining insight into the developmental mechanisms underlying fate choices in human pancreas and in developing cell-based therapies.
Stamcelinstituut Leuven, Laboratory for Experimental Medicine and Endocrinology, Department of Human Genetics, and University Hospitals Leuven, KU Leuven, Leuven, Belgium; Diabetes Research Center, Vrije Universiteit Brussel, Brussels, Belgium.
Qing
Q
Cai
Paola
P
Bonfanti
Rangarajan
R
Sambathkumar
Kim
K
Vanuytsel
Jolien
J
Vanhove
Conny
C
Gysemans
Maria
M
Debiec-Rychter
Susanna
S
Raitano
Harry
H
Heimberg
Laura
L
Ordovas
Catherine M
CM
Verfaillie
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000096:biosynthesis* / Q000235:genetics; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D064987:Cell- and Tissue-Based Therapy; D053379:Chromogranin A / Q000096:biosynthesis / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D005786:Gene Expression Regulation / Q000502:physiology*; D064593:Heterografts; D018398:Homeodomain Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000235:genetics; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics
D051792:Basic Helix-Loop-Helix Proteins; D053379:Chromogranin A; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D015534:Trans-Activators; C498695:NEUROG3 protein, human; C105201:NKX6-1 protein, human; C084010:pancreatic and duodenal homeobox 1 protein
Diabetes; Differentiation; Embryonic stem cells; Gene targeting; Progenitor cells