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Stem Cells International
Volume 2016, Article ID 6183562, 9 pages
Research Article

Dynamic Proteomic Analysis of Pancreatic Mesenchyme Reveals Novel Factors That Enhance Human Embryonic Stem Cell to Pancreatic Cell Differentiation

1Diabetes Center, Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA
2Department of Cell and Developmental Biology, Faculty of Medicine, Tel Aviv University, 69978 Tel Aviv, Israel
3Bioinformatics and High-Throughput Analysis Laboratory and High-Throughput Analysis Core, Center for Developmental Therapeutics, Seattle Children’s Research Institute, Seattle, WA 98105, USA
4Predictive Analytics, Seattle Children’s Hospital, Seattle, WA 98105, USA
5Data-Enabled Life Sciences Alliance (DELSA), Seattle, WA 98105, USA
6Department of Biomedical Informatics & Medical Education and Pediatrics, Medical School, University of Washington, Seattle, WA 98101, USA

Received 16 March 2015; Accepted 4 May 2015

Academic Editor: Stefan Liebau

Copyright © 2016 Holger A. Russ et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplementary Table 1: Spreadsheet containing all and GO term proteomic results.

Supplementary Figure 2S: A: RNA was isolated from bulk pancreatic tissues (total pancreas), pancreatic endothelial cells (Pecam1+ cells, isolated by FACS from total pancreata) and pancreatic mesenchymal cells (Nkx3.2/YFP+ cells, isolated by FACS from Nkx3.2-Cre;YFP total pancreata). Expression levels were analyzed by qPCR. N = 4. ***P < 0.005 as compared to total untreated pancreata. ND = Not detected. B: Treatment with LAMA2 partially inhibits culture-induced beta-cell dedifferentiation. RNA was extracted from freshly isolated islets (white) or from trypsin-dispersed islet cells after 3 days of culture on plates coated with either Poly-D-Lysine (blue) or human Merosin (a mixture of Laminin-211 and -221, green). N = 4. Data show one representative of two independent experiments with comparable results. *P < 0.05, **P < 0.01, ***P < 0.005, NS = non significant, as compared to freshly isolated islets.

Supplementary Figure 3S: Galectin-1 and α-2 chain Laminins are expressed only at very low levels after 12 days of differentiation of hES cells compared to human fibroblasts and islets while Neuroplastin is readily detectable. Quantitative PCR analysis of LGALS1, NPTN and LAMA2 transcripts in human foreskin fibroblasts (n=2) and purified human islet preparations (n=5) (as shown in figures 2&3) and hES after 12 days of differentiation (n=4, 2 independent experiments). Expression values are shown as average fold change ± standard deviation compared to expression levels of the endogenous control gene TBP.

  1. Supplementary Materials