TOP > 外国特許検索 > TECHNIQUE FOR ISOLATING/CULTURING COLORECTAL EPITHELIAL STEM CELL, AND TECHNIQUE FOR TRANSPLANTING COLORECTAL EPITHELIUM EMPLOYING SAID TECHNIQUE

TECHNIQUE FOR ISOLATING/CULTURING COLORECTAL EPITHELIAL STEM CELL, AND TECHNIQUE FOR TRANSPLANTING COLORECTAL EPITHELIUM EMPLOYING SAID TECHNIQUE

外国特許コード F130007364
整理番号 S2012-0054-N0
掲載日 2013年5月15日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2012JP006897
国際公開番号 WO 2013061608
国際出願日 平成24年10月26日(2012.10.26)
国際公開日 平成25年5月2日(2013.5.2)
優先権データ
  • 特願2011-236469 (2011.10.27) JP
発明の名称 (英語) TECHNIQUE FOR ISOLATING/CULTURING COLORECTAL EPITHELIAL STEM CELL, AND TECHNIQUE FOR TRANSPLANTING COLORECTAL EPITHELIUM EMPLOYING SAID TECHNIQUE
発明の概要(英語) The purpose of the present invention is to provide: a culture medium for the in vitro culture of a colorectal epithelial stem cell or the like; a method for culturing a colorectal epithelial stem cell or the like in vitro using the culture medium; a prophylactic/therapeutic agent for bowel diseases, which comprises a colorectal epithelial stem cell or the like produced by culturing by the aforementioned method; a method for administering, i.e., transplanting, a colorectal epithelial stem cell or the like produced by culturing by the aforementioned method to a patient suffering from a bowel disease; and a method for isolating a colorectal epithelial stem cell or the like. The culture medium, the prophylactic/therapeutic agent and the methods are characterized by using a culture medium for the in vitro culture of a colorectal epithelial stem cell and/or a colorectal epithelial cell, which comprises serum albumin, Wnt3a and r-spondin-1.
従来技術、競合技術の概要(英語) BACKGROUND ART
In the treatment of diseases in the digestive tract of the adult stem cells for possible therapeutic applications which are of interest. Stomach, small intestine and colon epithelial cells have the ability to generate the ability to self-replicate and gastrointestinal epithelial stem cells, which are parts of the digestive tract play an important role in the maintenance of tissue homeostasis (non-patent document 1-3) play. A recent study, stem cells of the gastrointestinal tract is largely increased biological findings are. Wnt, some BMP Notch and a cascade of signal transduction pathways and are simultaneously applied, adjusting the maintenance of stem cells, epithelial homeostasis control (non-patent document 4) are shown. Studies using the tracking system, the target gene is Wnt Lgr5, small intestine and colon crypts (non-patent document 5) as well as the bottom of the stomach (non-patent document 6) unit at the bottom of which is identified as a marker of stem cells.
Current, reliable markers can be recognized by stem cells from the gastrointestinal tract, the gastrointestinal tract many cells with stem cell therapy for the treatment of diseases refractory to fundamentally change being expected. However to this day, gastrointestinal epithelial cells isolated and cultured in vitro after incrementing the, cell therapy to repair the injured tissue used as the material of there is no report. In addition, culture and propagate gastrointestinal epithelial stem cell injury can be restored to a tissue at the site as yet not been found. Mesenchymal and epithelial component a small fragment of an intestinal component over a long period of culture can be a recently (non-patent document 7).
Non-patent document 8 is, colon crypt was isolated and embedded in the matrigel, the destination completely negative fossa culture added, the method comprising culturing a colon epithelial stem cells is described. A completely negative fossa culture point that is, Advanced DMEM/F12 to a basal medium, conditioned medium Wnt3a or Wnt3a, EGF, Noggin, r-spondin-1, (registered trademark) supplement(Invitrogen Corporation) B-27, gastrin, nicotinamide, A83-01 (TGF-β receptor kinase inhibitor 1), and, such as SB202190 (p38 inhibitor) which is a medium was added. (Registered trademark) supplement B-27 precise composition is not disclosed, biotin, carnitine L -, corticosterone, ethanolamine, D (+) galactose, reduced glutathione, linoleic acid, linolenic acid, progesterone, putrescine, retinyl acetate, selenium, -l- chiromin toriodo, vitamin E, vitamin E acetate, bovine albumin, catalase, insulin, superoxide, dismutase, and the like known transferrin (non-patent document 9). Colon epithelial stem cells cultured in vitro in a serum-free medium, gastrin, nicotinamide, A83-01 (TGF-β receptor kinase inhibitor 1), SB202190 (p38 inhibitor), and, an essential component B-27 supplement are believed to, among, and is particularly essential B-27 supplement has been considered.
However, bovine serum albumin (bovine serum albumin; BSA) is, contained a lot in the bovine serum albumin, fatty acids and trace elements function as carriers has been known. Is Wnt3a, and the protein encoded by the gene WNT3A, β - catenin pathways known to activate. - Catenin pathway is Wnt / β -, or proliferation of the stem cell, is related to a maintenance anaplasia is known. R-spondin-1(Roof-plate-specific Spondin-1; Rspo1) is, and the protein encoded by the gene RSPO1, contribute to the differentiation of dorsal neural tube, of promoting proliferation of endothelial cells crypt intestinal anion is known. Epidermal growth factor (Epidermal Growth Factor; EGF) is, and the protein encoded by the gene EGF, promote proliferation of epithelial cells has been known. Hepatocyte growth factor (hepatocyte growth factor; HGF) is, HGF protein encoded by the gene, as well as a hepatocytes, promoting the growth of various cells are known. Noggin is, and the protein encoded by the NOG gene, transforming growth factor β (Transforming growth factor beta; TGF-β) signaling is known to inhibit.
However, the basic medium components (sugars of a carbon source, nitrogen source such as amino acids, inorganic salts) in addition to, serum albumin, Wnt3a, and, using r-spondin-1, culture in serum-free medium, a mammal such as a mouse or a human colon isolated from tissue such as colon epithelial stem cells in vitro for long periods of time, may amplify has not been known at all.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • NATIONAL UNIVERSITY CORPOPRATION TOKYO MEDICAL AND DENTAL UNIVERSITY
  • 発明者(英語)
  • WATANABE, Mamoru
  • NAKAMURA, Tetsuya
国際特許分類(IPC)
指定国 National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IS JP KE KG KM KN KP KR KZ LA LC LK LR LS LT LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG
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