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Endocrine Cell Clustering During Human Pancreas Development

Jongmin Jeon¹, Mayrin Correa-Medina¹, Camillo Ricordi, Helena Edlund and Juan A. Diez

Diabetes Research Institute, University of Miami Leonard M. Miller School of Medicine, Miami, Florida (JJ,MC-M,CR,JAD), and Umeå Center for Molecular Medicine, Umeå University, Umeå, Sweden (HE)

Correspondence to: Juan Diez, Diabetes Research Institute, University of Miami Leonard M. Miller School of Medicine, 1450 NW 10th Avenue (R-134), Miami, FL 33136. E-mail: jdiez{at}med.miami.edu

The development of efficient, reproducible protocols for directed in vitro differentiation of human embryonic stem (hES) cells into insulin-producing β cells will benefit greatly from increased knowledge regarding the spatiotemporal expression profile of key instructive factors involved in human endocrine cell generation. Human fetal pancreases 7 to 21 weeks of gestational age, were collected following consent immediately after pregnancy termination and processed for immunostaining, in situ hybridization, and real-time RT-PCR expression analyses. Islet-like structures appear from approximately week 12 and, unlike the mixed architecture observed in adult islets, fetal islets are initially formed predominantly by aggregated insulin- or glucagon-expressing cells. The period studied (7–22 weeks) coincides with a decrease in the proliferation and an increase in the differentiation of the progenitor cells, the initiation of NGN3 expression, and the appearance of differentiated endocrine cells. The present study provides a detailed characterization of islet formation and expression profiles of key intrinsic and extrinsic factors during human pancreas development. This information is beneficial for the development of efficient protocols that will allow guided in vitro differentiation of hES cells into insulin-producing cells. (J Histochem Cytochem 57:811–824, 2009)

Key Words: fetal pancreas • gene expression profile • human • immunohistochemistry • islet formation

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