title: Mapping the Cell-Surface N-Glycoproteome of Human Hepatocytes Reveals Markers for Selecting a Homogeneous Population of iPSC-Derived Hepatocytes.
abstract: When comparing hepatic phenotypes between iPSC-derived hepatocyte-like cells from different liver disease patients, cell heterogeneity can confound interpretation. We proposed that homogeneous cell populations could be generated by fluorescence-activated cell sorting (FACS). Using cell-surface capture proteomics, we identified a total of 300 glycoproteins on hepatocytes. Analyses of the expression profiles during the differentiation of iPSCs revealed that SLC10A1, CLRN3, and AADAC were highly enriched during the final stages of hepatocyte differentiation. FACS purification of hepatocyte-like cells expressing SLC10A1, CLRN3, or AADAC demonstrated enrichment of cells with hepatocyte characteristics. Moreover, transcriptome analyses revealed that cells expressing the liver gene regulatory network were enriched while cells expressing a pluripotent stem cell network were depleted. In conclusion, we report an extensive catalog of cell-surface N-linked glycoproteins expressed in primary hepatocytes and identify cell-surface proteins that facilitate the purification of homogeneous populations of iPSC-derived hepatocyte-like cells.
authors: Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29425, USA.
authors: Sunil K
authors: SK
authors: Mallanna
authors: Department of Cell Biology, Neurobiology and Anatomy, Program in Regenerative Medicine, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
authors: Max A
authors: MA
authors: Cayo
authors: Department of Cell Biology, Neurobiology and Anatomy, Program in Regenerative Medicine, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Department of Pediatrics, University of Minnesota, Minneapolis, MN 55454, USA.
authors: Kirk
authors: K
authors: Twaroski
authors: Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
authors: Rebekah L
authors: RL
authors: Gundry
authors: Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29425, USA. Electronic address: duncanst@musc.edu.
authors: Stephen A
authors: SA
authors: Duncan
mesh_terms: D015415:Biomarkers; D002454:Cell Differentiation; D016000:Cluster Analysis; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D009928:Organ Specificity / Q000235:genetics; D010641:Phenotype; D020543:Proteome; D040901:Proteomics / Q000379:methods
chemical_list: D015415:Biomarkers; D008562:Membrane Glycoproteins; D020543:Proteome