Research Article

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

Figure 4

Mesenchymal factors, Laminins containing a α-2 chain and Galectin-1, improve human embryonic stem cell differentiation. (a) Schematic of the experimental setup to test possible effects of novel mesenchymal factors on direct differentiation of human embryonic stem (ES) cells. Factors were added at day 9 to posterior foregut-like (PF) cells for three days before analysis of pancreatic progenitor (PP) cells. (b) Flow cytometric analysis of proliferating cells in control and treated cultures at day 12 after 24-hour incubation with BrdU. Data shown are from 2 independent experiments, . (c) Quantitative PCR analysis for the pancreatic progenitor marker PDX1 and endocrine marker NGN3. Data shown are average ± standard deviation from 3 independent experiments, , relative to untreated control cultures at day 12 of differentiation and normalized to the endogenous control gene TBP. (d) Quantification of immunofluorescence analysis of PDX1 positive as a percentage of total cells with an InCell Analyzer. Data shown are average ± standard deviation of 16 independent fields assayed in 2 independent experiments. Size bar = 250 μm.