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.
Bibliographical noteFunding Information:
The authors acknowledge Hope Campbell at Blood Center of Wisconsin for performing FACS sorting. This work was supported by gifts from the Marcus Family , the Phoebe R. and John D. Lewis Foundation , the Sophia Wolf Quadracci Memorial Fund , the Advancing a Healthier Wisconsin Fund , and by NIH grants DK102716 , HG006398 , and HD082570 to S.A.D. and NIH grant HL094708 to R.L.G. Confocal analyses were supported by COBRE P30 GM103342 . We would like to dedicate this manuscript to the memory of John (Jack) Lewis (1927–2015).
© 2016 The Author(s)