title: Generation of insulin-producing β-like cells from human iPS cells in a defined and completely xeno-free culture system.
abstract: Human induced pluripotent stem (hiPS) cells are considered a potential source for the generation of insulin-producing pancreatic β-cells because of their differentiation capacity. In this study, we have developed a five-step xeno-free culture system to efficiently differentiate hiPS cells into insulin-producing cells in vitro. We found that a high NOGGIN concentration is crucial for specifically inducing the differentiation first into pancreatic and duodenal homeobox-1 (PDX1)-positive pancreatic progenitors and then into neurogenin 3 (NGN3)-expressing pancreatic endocrine progenitors, while suppressing the differentiation into hepatic or intestinal cells. We also found that a combination of 3-isobutyl-1-methylxanthine (IBMX), exendin-4, and nicotinamide was important for the differentiation into insulin single-positive cells that expressed various pancreatic β-cell markers. Most notably, the differentiated cells contained endogenous C-peptide pools that were released in response to various insulin secretagogues and high levels of glucose. Therefore, our results demonstrate the feasibility of generating hiPS-derived pancreatic β-cells under xeno-free conditions and highlight their potential to treat patients with type 1 diabetes.
authors: Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo 2-2-1, Chuo-Ku, Kumamoto 860-0811, Japan Global-Center of Excellence (G-COE), Kumamoto University, Honjo 2-2-1, Chuo-ku, Kumamoto 860-0811, Japan.
authors: Hussain Md
authors: HM
authors: Shahjalal
authors: Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo 2-2-1, Chuo-Ku, Kumamoto 860-0811, Japan.
authors: Nobuaki
authors: N
authors: Shiraki
authors: Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo 2-2-1, Chuo-Ku, Kumamoto 860-0811, Japan Global-Center of Excellence (G-COE), Kumamoto University, Honjo 2-2-1, Chuo-ku, Kumamoto 860-0811, Japan.
authors: Daisuke
authors: D
authors: Sakano
authors: Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo 2-2-1, Chuo-Ku, Kumamoto 860-0811, Japan Global-Center of Excellence (G-COE), Kumamoto University, Honjo 2-2-1, Chuo-ku, Kumamoto 860-0811, Japan Department of Pediatrics, Graduate School of Medical Sciences, Kumamoto University, Honjo 1-1-1, Chuo-Ku, Kumamoto 860-8556, Japan.
authors: Kazuhide
authors: K
authors: Kikawa
authors: Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo 2-2-1, Chuo-Ku, Kumamoto 860-0811, Japan.
authors: Soichiro
authors: S
authors: Ogaki
authors: Department of Gastroenterological Surgery, Graduate School of Medical Sciences, Kumamoto University, Honjo 1-1-1, Chuo-Ku, Kumamoto 860-8556, Japan.
authors: Hideo
authors: H
authors: Baba
authors: Department of Neuropharmacology, Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe Street, Mizuho, Nagoya 467-8603, Japan.
authors: Kazuhiko
authors: K
authors: Kume
authors: Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo 2-2-1, Chuo-Ku, Kumamoto 860-0811, Japan Global-Center of Excellence (G-COE), Kumamoto University, Honjo 2-2-1, Chuo-ku, Kumamoto 860-0811, Japan Program for Leading Graduate Schools 'HIGO (Health Life Science; Interdisciplinary and Glocal Oriented) Program,' Kumamoto University, Honjo 2-2-1, Chuo-ku, Kumamoto 860-0811, Japan skume@kumamoto-u.ac.jp.
authors: Shoen
authors: S
authors: Kume
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D003470:Culture Media; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007328:Insulin / Q000096:biosynthesis; D050417:Insulin-Secreting Cells / Q000166:cytology*
chemical_list: D003470:Culture Media; D007328:Insulin
keywords: cell therapy; diabetes; hiPS cells; pancreas; xeno-free differentiation; β-cells