In conclusion the data presented here support

In conclusion, the data presented here support a novel strategy to generate pancreatic β-like cells. The gall mCAP is a readily accessible and non-essential tissue that contains cells amenable to large-scale expansion and reprogramming to a pancreatic fate. In addition, our data indicate that this endodermal derivative is more amenable to reprogramming than skin fibroblasts. However, the reprogramming is currently only partial and the rGBCs did not become fully functional, mature β-cells in vitro. Although there were many up-regulated marker genes detected in rGBCs, global RNA-Seq analysis was needed to properly characterize the cells relative to true functional β-cells, something that is commonly missing in other reprogramming studies. Nonetheless, the results outlined here will be useful for further experiments aimed at generating a direct differentiation-based cell therapy for type 1 diabetes in human patients. The following are the supplementary data related to this article.
Acknowledgments We thank Pamela Canaday (Flow Cytometry Resource at OHSU) for cell sorting. We thank Jessie Coleman for technical support with the mouse colony. We also thank Milton Finegold and Angela Major (NIDDK-sponsored Digestive Disease Core Laboratory of the Texas Medical Center (DK56338)) for histology support and to Elisabetta Manduchi (UPenn) for helping to transfer data. We are grateful to Craig Dorrell (OHSU) for thoughtful discussions. This work was supported by grants from NIH/NIDDK grants U01 DK072477 (MG), NIH grants AI46629 (DLG, LDS) and DK89572 (DLG, LDS), an institutional Diabetes Endocrinology Research Center (DERC) grant DK32520 (DLG, MAB, LDS), grants from the Helmsley Foundation (MG, DLG, LDS) and the Juvenile Diabetes Research Fund (MG).
Introduction The adult forebrain contains neural stem cells (NSCs) within the subventricular zone (SVZ), and these cells ensure neurogenesis during adulthood (Doetsch et al., 1999a). Adult NSCs successively give rise to transit amplifying cells (TACs) and then neuroblasts. Neuroblasts then migrate in chains to the olfactory bulbs, where they differentiate into neurons (Lois et al., 1996). Adult-born neurons in the olfactory bulbs are critical for olfactory memory (Lazarini et al., 2009). Adult NSCs, also known as type B cells, contact the lateral ventricle, have an astrocytic phenotype and express Glial Fibrillary Acidic Protein (GFAP) (Doetsch et al., 1999a; Garcia et al., 2004). NSCs also express the cell surface carbohydrate Lewis X (LeX)/CD15 (Capela and Temple, 2002; Obermair et al., 2010). Adult NSCs have been considered relatively quiescent, with a cell cycle length lasting several days under physiological conditions, in contrast to 12h for TACs (Morshead et al., 1994). Killing the constitutively proliferating cells in the SVZ using antimitotic drugs leaves quiescent NSCs intact, has few effects on the number of NSCs and results in a complete recovery of neurogenesis in the SVZ through repopulation from the relatively quiescent NSCs (Doetsch et al., 1999b; Morshead et al., 1994). Similarly, exposure of the adult brain to a moderate dose of ionizing radiation provokes the death of proliferating cells, such as TACs and neuroblasts (Chevalier et al., 2012; Shinohara et al., 1997; Tada et al., 1999). This phenomenon is followed by NSC proliferation and the recovery of cells in the SVZ, suggesting that quiescent NSCs are activated to re-enter the cell cycle with major alterations in oxidative metabolism (Chevalier et al., 2012). Using GFAP::GFP transgenic mice combined with CD24 and a fluorophore-complexed EGF ligand, Pastrana et al. reported the appearance of cycling NSCs with a GFAP+EGFR+ phenotype after exposure to an antimitotic drug; using a FACS approach, the authors prospectively isolated these activated NSCs (Pastrana et al., 2009). These findings suggest that a population of quiescent NSCs exists that are capable of re-entering the cell cycle and restoring neurogenesis after SVZ depletion. However, recent studies fail to show that quiescent adult NSCs exist outside the cell cycle and remain in stasis until activated by appropriate proliferative signals, and to-date, no prospective methods have been reported to sort this population of cells.