title: Regeneration of insulin-producing pancreatic cells using a volatile bioactive compound and human teeth.
abstract: Transplantation of insulin (INS)-secreting cells differentiated in vitro from stem cells promises a safer and easier treatment of severe diabetes mellitus. A volatile bioactive compound, hydrogen sulfide (H2S), promotes cell differentiation; human tooth-pulp stem cells undergo hepatic differentiation. The aim of this study is to develop a novel protocol using H2S to enhance pancreatic differentiation from the CD117(+) cell fraction of human tooth pulp. During the differentiation, the cells were exposed to 0.1 ng ml(-1) H2S. Immunocytochemistry, RT-PCR, determination of INS c-peptide content and flow cytometry of pancreatically related markers were carried out. Expression of WNT and the PI3K/AKT signaling pathway were also determined by PCR array. After differentiation, INS, glucagon (GCG), somatostatin (SST) and pancreatic polypeptide (PPY) were positive when examined by immunofluorescence. INS and GCG were also determined flow-cytometrically. Only the cells expressing INS increased after H2S exposure. The number of cells expressing GCG was significantly decreased. Genes involved in canonical WNT and the WNT/calcium pathways were highly expressed after H2S exposure. H2S accelerated INS synthesis and secretion by regenerated INS-producing cells from human teeth. All signaling pathway functions of the PI3K-AKT pathway were extremely activated by H2S exposure. The matured INS-producing cells originating in human teeth were increased by H2S in order to control blood-glucose level.
authors.affiliation: Nippon Dental University, School of Life Dentistry at Tokyo, Department of Oral Health, 1-9-20 Chiyoda-ku, 102-8159 Tokyo, Japan.
authors.forename: Mio
authors.initials: M
authors.lastname: Okada
authors.forename: Toshio
authors.initials: T
authors.lastname: Imai
authors.forename: Ken
authors.initials: K
authors.lastname: Yaegaki
authors.forename: Nikolay
authors.initials: N
authors.lastname: Ishkitiev
authors.forename: Tomoko
authors.initials: T
authors.lastname: Tanaka
mesh_terms: D002096:C-Peptide / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D006862:Hydrogen Sulfide / Q000494:pharmacology; D007150:Immunohistochemistry; D007328:Insulin / Q000096:biosynthesis* / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012038:Regeneration / Q000187:drug effects*; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects; D014070:Tooth / Q000166:cytology*; D055549:Volatile Organic Compounds / Q000494:pharmacology*; D060449:Wnt Signaling Pathway / Q000187:drug effects / Q000235:genetics
chemical_list: D002096:C-Peptide; D007328:Insulin; D055549:Volatile Organic Compounds; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D005947:Glucose; D006862:Hydrogen Sulfide