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A Quick and Efficient Method for the Purification of Endoderm Cells Generated from Human Embryonic Stem Cells
Published on: March 3, 2016
Targeting SOX17 in human embryonic stem cells creates unique strategies for isolating and analyzing developing
Pei Wang1, Ryan T Rodriguez, Jing Wang
1Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Cell Stem Cell
|March 3, 2011
Summary
Researchers developed a new method using SOX17 gene tagging in human embryonic stem cells (hESCs) to isolate and study endodermal cells. This advance aids understanding of human embryonic endoderm development and differentiation.
Area of Science:
- Developmental Biology
- Stem Cell Research
- Genetics
Background:
- Human embryonic stem cells (hESCs) offer a model for studying inaccessible embryonic tissues like endoderm.
- Previous research faced challenges in isolating endodermal cells and tracking their developmental fates.
Purpose of the Study:
- To develop methods for isolating and tracing the differentiation of human endodermal cells derived from hESCs.
- To identify reliable markers for endodermal cell populations.
Main Methods:
- Homologous recombination was used to insert an enhanced green fluorescent protein (eGFP) transgene into the SOX17 locus of hESCs.
- Fluorescence-activated cell sorting (FACS) and gene expression profiling were employed for cell purification and analysis.
Main Results:
- SOX17-expressing cells derived from hESCs exhibited endodermal markers.
- Specific cell surface protein combinations were identified, enabling FACS-based isolation of primitive gut tube endodermal cells from hESCs and induced pluripotent stem cells (iPSCs).
- Differentiated SOX17(+) endodermal cells showed markers for liver, pancreas, and intestinal epithelium in vitro and formed endodermal progeny in vivo.
Conclusions:
- Prospective isolation and lineage tracing of SOX17(+) hESC progeny provide fundamental insights into human endodermal biology.
- This method facilitates developmental studies of endodermal lineages.
- The identified markers enable efficient isolation of specific endodermal cell types.
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