A molecular signature for purified definitive endoderm guides differentiation and isolation of endoderm from mouse and human embryonic stem cells

Pei Wang, Kristen D. McKnight, David J. Wong, Ryan T. Rodriguez, Takuya Sugiyama, Xueying Gu, Amar Ghodasara, Kun Qu, Howard Y. Chang, Seung K. Kim

Resultado de la investigación: Articlerevisión exhaustiva

30 Citas (Scopus)

Resumen

Embryonic definitive endoderm (DE) generates the epithelial compartment of vital organs such as liver, pancreas, and intestine. However, purification of DE in mammals has not been achieved, limiting the molecular "definition" of endoderm, and hindering our understanding of DE development and attempts to produce endoderm from sources such as embryonic stem (ES) cells. Here, we describe purification of mouse DE using fluorescence-activated cell sorting (FACS) and mice harboring a transgene encoding enhanced green fluorescent protein (eGFP) inserted into the Sox17 locus, which is expressed in the embryonic endoderm. Comparison of patterns of signaling pathway activation in native mouse DE and endoderm-like cells generated from ES cells produced novel culture modifications that generated Sox17-eGFP+ progeny whose gene expression resembled DE more closely than achieved with standard methods. These studies also produced new FACS methods for purifying DE from nontransgenic mice and mouse ES cell cultures. Parallel studies of a new human SOX17-eGFP ES cell line allowed analysis of endoderm differentiation in vitro, leading to culture modifications that enhanced expression of an endoderm-like signature. This work should accelerate our understanding of mechanisms regulating DE development in mice and humans, and guide further use of ES cells for tissue replacement.

Idioma originalEnglish (US)
Páginas (desde-hasta)2273-2287
Número de páginas15
PublicaciónStem Cells and Development
Volumen21
N.º12
DOI
EstadoPublished - ago. 10 2012
Publicado de forma externa

ASJC Scopus subject areas

  • Hematology
  • Developmental Biology
  • Cell Biology

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