|Cancer||Cell death||Cell cycle||Cytoskeleton||Exo/endocytosis||Differentiation||Division||Organelles||Signalling||Stem cells||Trafficking|
Identification of microRNAs expressed highly in pancreatic islet-like cell clusters differentiated from human embryonic stem cells
Bo‑Zhi Chen*1, Sung‑Liang Yu†1, Sher Singh‡, Li‑Pin Kao*, Zong‑Yun Tsai§║, Pan‑Chyr Yang¶, Bai‑Hsiun Chen* and Steven Shoei‑Lung Li*║2
*Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan, †Department of Clinical Laboratory Sciences and Medical Biotechnology, National Taiwan University College of Medicine, Taipei 100, Taiwan, ‡Department of Life Sciences, College of Science, National Taiwan Normal University, Taipei 116, Taiwan, §Department of Medicinal and Applied Chemistry, College of Life Sciences, Kaohsiung Medical University, Kaohsiung 807, Taiwan, ║Stem Cell Laboratory, Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan, and ¶Department of Internal Medicine, National Taiwan University College of Medicine, Taipei 100, Taiwan
Type 1 diabetes is an autoimmune destruction of pancreatic islet beta cell disease, making it important to find a new alternative source of the islet beta cells to replace the damaged cells. hES (human embryonic stem) cells possess unlimited self-renewal and pluripotency and thus have the potential to provide an unlimited supply of different cell types for tissue replacement. The hES-T3 cells with normal female karyotype were first differentiated into EBs (embryoid bodies) and then induced to generate the T3pi (pancreatic islet-like cell clusters derived from T3 cells), which expressed pancreatic islet cell-specific markers of insulin, glucagon and somatostatin. The expression profiles of microRNAs and mRNAs from the T3pi were analysed and compared with those of undifferentiated hES-T3 cells and differentiated EBs. MicroRNAs negatively regulate the expression of protein-coding mRNAs. The T3pi showed very high expression of microRNAs, miR-186, miR-199a and miR-339, which down-regulated the expression of LIN28, PRDM1, CALB1, GCNT2, RBM47, PLEKHH1, RBPMS2 and PAK6. Therefore, these microRNAs and their target genes are very likely to play important regulatory roles in the development of pancreas and/or differentiation of islet cells, and they may be manipulated to increase the proportion of beta cells and insulin synthesis in the differentiated T3pi for cell therapy of type I diabetics.
Key words: expression profile, human embryonic stem cell, microRNA, mRNA, pancreatic islet-like cell, target identification
Abbreviations: bFGF, basic fibroblast growth factor; DMEM, Dulbecco's modified Eagle's medium, EBs, embryoid bodies, GSEA, MetaCore Gene Set Enrichment Analysis, hES, human embryonic stem, ITSF, insulin, transferrin, selenium, fibronectin, MEF, mouse embryonic fibroblast, miRNAs, microRNAs, NGN3, neurogenin 3, PDX1, pancreatic duodenal homeobox 1; RT, reverse transcription, T3EB, embryoid bodies differentiated from T3 cells, T3ES, hES-T3 cells grown on MEF feeder, T3pi, pancreatic islet-like cell clusters derived from T3 cells
1Bo-Zhi Chen and Sung-Liang Yu contributed equally to this work.
2To whom correspondence should be addressed (email firstname.lastname@example.org).
The original data of T3pi cells obtained from Affymetrix human genome U133 plus 2.0 GeneChip have been deposited to NCBI database, and the GEO series number is GSE14503.
Received 3 July 2009/2 December 2009; accepted 25 August 2010
Published as Cell Biology International Immediate Publication 25 August 2010, doi:10.1042/CBI20090081
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