Stem cell regulation towards cell therapy for diabetes

• Project/Area Number: 17209043
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General surgery
• Research Institution: Yokohama City University
• Principal Investigator: TANIGUCHI Hideki Yokohama City University, Graduate School of Medicine Department of Regenerative Medicine, Prof (70292555)
• Project Period (FY): 2005 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥49,530,000 (Direct Cost: ¥38,100,000、Indirect Cost: ¥11,430,000); Fiscal Year 2007: ¥14,430,000 (Direct Cost: ¥11,100,000、Indirect Cost: ¥3,330,000); Fiscal Year 2006: ¥14,430,000 (Direct Cost: ¥11,100,000、Indirect Cost: ¥3,330,000); Fiscal Year 2005: ¥20,670,000 (Direct Cost: ¥15,900,000、Indirect Cost: ¥4,770,000)
• Keywords: Stem Cell / Diabetes / Pancreatic islet / Regeneration

Research Abstract

Islet transplantation has become available across the globe since a novel protocol was reported. However, because donors are in short supply, only a minority of patients benefit from this procedure. Pancreatic progenitor cells are a promising resource for regeneration of new islets but if progenitor cells reside in ductal epithelium is not clear. Mouse pancreas was examined by immunohistochemistry with cell surface markers specific for ductal cells. We developed an isolation method for ductal cells by flow cytometric cell sorting using a newly identified specific marker for ductal cells. Using an in vitro colony assay, we characterized their proliferative and multipotent capacity. CD133 is specifically expressed in ductal epithelium. Flow cytometric analysis revealed that purified ductal cells are highly enriched in the CD133^+CD34^-CD45^-Ter119^- fraction. Analysis of clonal epithelial colonies formed by individual cells revealed that progenitor cells with multilineage differentiation capacity are present in neonatal ductal epithelium. Moreover, these progenitor cells express c-Met. In adult mice, progenitor cells that exhibit a high proliferative capacity but appear committed to a ductal lineage are co-purified with CD133^+CD34^-CD45^-Ter119^-cells. We established a system for isolating and culturing mouse pancreatic ductal cells that relies on flow cytometric cell sorting. Clonal analysis revealed that a population of progenitor cells is present amongst CD133^+ ductal cells. Isolation of these cells will facilitate future studies into the roles of pancreatic progenitor cells in regeneration and carcinogenesis.

Research Products

• [Journal Article] Tbx3 controls the fate of hepatic progenitor cells in development by suppressing p19ARF expression (2008)
  • Author(s): Suzuki A, Sekiya S, Buscher D, Izpisua Belmonte JC, Taniguchi H
  • Journal Title: Decelopment 135 Pages: 1589-1595
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Tbx3 controls the fate of hepatic progenitor cells in liver development by suppressing p19ARF expression (2008)
  • Author(s): Suzuki, A., Sekiya, S., Buscher, D., Izpisua, Belmonte, JC, Taniguchi, H
  • Journal Title: Development 135(9) Pages: 1589-1595
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Tbx3 controls the fate of hepatic progenitor cells in liver development by suppressing p19ARF expression (2008)
  • Author(s): Suzuki A, Sekiya S, Buscher D, Izpisua Belmonte JC, Taniguchi H
  • Journal Title: Development 135 Pages: 1589-1595
  • Peer Reviewed
• [Journal Article] Enhanced self-renewal capability in hepatic stem/progenitor cells drives cancer initiation (2007)
  • Author(s): Chiba T, Zhen YW, Kita K, Yokosuka O, Saisho H, Onodera M, Miyoshi H, Nakano M, Zen Y, Nakanuma Y, Nakauchi H, Iwama A, Taniguchi H
  • Journal Title: Gastroenterology 133 Pages: 937-950
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Adult rat hepatic bipotent progenitor cels remain doramant even after extensive hepatectomy (2007)
  • Author(s): Yamamoto Y, Togo S, Yun-wen Zheng, Kubota T, Taniguchi H, Shimada S
  • Journal Title: Sound Repair and Regeneration 15 Pages: 422-429
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Neonatal pancreati ccells redifferentiate into both neural and pancreatic lineages (2007)
  • Author(s): Zhao W
  • Journal Title: Biochemical and Biophysical Research Communications 352(1) Pages: 84-90
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] In vitro reconstitution of hepatic tissue architectures with neonatal mouse liver cells usingt hree-dimensional culture (2007)
  • Author(s): Okamura A
  • Journal Title: Journal of Nanoscience and Nanotechnology 7(3) Pages: 721-725
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Isolation of mouse pancreatic ductal progenitor cells expressing CD133 and c-Met by flow cytometxi ccell sorting. (2007)
  • Author(s): Oshima Y
  • Journal Title: Gastroenterology 132(2) Pages: 720-732
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Production of Functional Spermatids from Mouse Germline Stem Cells in Ectopically Reconstituted Seminiferous Tubules (2007)
  • Author(s): Kita K
  • Journal Title: Biol Reprod. 76(2) Pages: 211-217
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Enhanced self-renewal capability in hepatic stem/progenitor cells drives cancer initiation (2007)
  • Author(s): Chiba, T., Zhen, Y.W., Kita, K., Yokosuka, O., Saisho, H., Onodera, M., Miyoshi, H., Nakano, M., Zen, Y., Nakanuma, Y., Nakauchi, H., Iwama, A., Taniguchi, H
  • Journal Title: Gastroenterology 133(3) Pages: 937-950
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Adult rat hepatic bipotent progenitor cels remain doramant even after extensive hepatectomy (2007)
  • Author(s): Yamamoto, Y., Togo, S., Yun-wen, Zheng, Kubota, T., Taniguchi, H., Shimada, S
  • Journal Title: Wound Repair and Regeneration 15(3) Pages: 422-429
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Neonatal pahcreatic cells redifferentiate into both neural and Communications (2007)
  • Author(s): Zhao, W., Hirose, T., Ishikawa, M., Oshima, Y., Hirai, S., Ohno, S., Taniguchi, H
  • Journal Title: Biochemical and Biophysical Research 352(1) Pages: 84-90
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] In vitro reconstitution of hepatic tissue architectures with neonatal mouse liver cells using three-dimensional culture (2007)
  • Author(s): Okamura, A., Zheng, Y.W., Hirochika, R., Tanaka, J., Taniguchi, H
  • Journal Title: Journal of Nanoscience and Nanotechnology 7(2) Pages: 721-5
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Isolation of mouse pancreatic ductal progenitor cells expressing CD133 and c-Met by flow cytometric cell sorting (2007)
  • Author(s): Oshima, Y., Suzuki, A., Kawashimo, K., Ishikawa, M., Ohkohchi, N., Taniguchi, H
  • Journal Title: Gastroenterology 132(2) Pages: 720-732
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Production of Functional Spermatids from Mouse Germline Stem Cells in Ectopically Reconstituted Seminiferous Tubules (2007)
  • Author(s): Kita, K., Watanabe, T., Ohsaka, K., Hayashi, H., Kubota, Y., Nagashima, Y., Aoki, I., Taniguchi, H., Noce, T., Inoue, K., Miki, H., Ogonuki, N., Tanaka, H., Ogura, A., Ogawa, T
  • Journal Title: Biol Reprod 76(2) Pages: 211-7
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Enhanced self-renewal capability in hepatic stem/progenitor cells drives cancer initiation (2007)
  • Author(s): Chiba T, Zhen YW, Kita K, Yokosuka O, Saisho H, Onodera M, Miyoshi H, Nakano M, Zen Y, Nakanuma Y, Nakauchi H, Iwama A, Taniuchi H
  • Journal Title: Gastroenterology 133 Pages: 937-950
  • Peer Reviewed
• [Journal Article] Neonatal pancreatic cells redifferentiate into both neural and pancreatic lineages (2007)
  • Author(s): Zhao W
  • Journal Title: Biochemical and Biophysical Research Communications 352(1)
• [Journal Article] In vitro reconstitution of hepatic tissue architectures with neonatal mouse liver cells using three-dimensional culture (2007)
  • Author(s): Okamura A
  • Journal Title: Journal of Nanoscience and Nanotechnologv 7(3) Pages: 721-725
• [Journal Article] Isolation of mouse pancreatic ductal progenitor cells expressing CD 133 and c-Met by flow cytometric cell sorting. (2007)
  • Author(s): Oshima Y
  • Journal Title: Gastroenterology 132(2) Pages: 720-732
• [Journal Article] Clonal proliferation of multipotent stem/progenitor cells in the neonatal and adultsalivary glands (2006)
  • Author(s): Kishi T
  • Journal Title: Biochemical and Biophysical Research Communications 340 Pages: 544-552
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Side Population Purified From Hepatocellular Carcinoma Cells Harbors Cancer Stem Cell-like Properties (2006)
  • Author(s): Chiba T
  • Journal Title: Hepatology 44(1) Pages: 240-251
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Clonal proliferation of multipotent stem/progenitor cells in the neonatal and adult salivary glands (2006)
  • Author(s): Kishi, T., Takao, T., Fujita, K., Taniguchi, H
  • Journal Title: Biochemical and Biophysical Research Communications 340 Pages: 544-552
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Side Population Purified From Hepatocellular Carcinoma Cells Harbors Cancer Stem Cell- like Properties (2006)
  • Author(s): Chiba, T., Kita, K., Zhen, Y.W., Yokosuka, O., Saisho, H., Iwama, A., Nakauchi, H., Tanisuchi, H
  • Journal Title: Hepatology 44(1) Pages: 240-251
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] 膵臓における組織幹細胞の分離・同定 (2006)
  • Author(s): 谷口英樹
  • Journal Title: 最新医学 61(7) Pages: 28-34
• [Journal Article] Clonal proliferation of multipotent stem/progenitor cells in the neonatal and adult salivary glands (2006)
  • Author(s): Kishi T
  • Journal Title: Biochemical and Biophysical Research Communications 340 Pages: 544-552
• [Journal Article] 肝臓・胆管・膵臓の発生学的な関連性 (2006)
  • Author(s): 谷口英樹
  • Journal Title: 肝胆膵 52(2) Pages: 167-172
• [Journal Article] 肝臓の再生は?「肝臓を再生することはできますか?」 (2005)
  • Author(s): 谷口英樹
  • Journal Title: Q&Aでわかる肥満と糖尿病 特集 肝臓--脂肪性肝炎とC型肝炎-- (5)(6) Pages: 1079-1081
• [Journal Article] 糖尿病の再生医学-ES細胞研究の現状と課題 (2005)
  • Author(s): 谷口英樹
  • Journal Title: 再生医療 2(4-1) Pages: 135-140
• [Journal Article] 肝臓における組織幹細胞の分離と特性解析 (2005)
  • Author(s): 谷口英樹
  • Journal Title: 総合臨床 54(1) Pages: 107-112
• [Presentation] Excessive Self-renewal of Hepatic Stem Cells Drives Cancer Initiation (2008)
  • Author(s): Taniguchi, H
  • Organizer: International Symposium on Tumor Biology in Kanazawa 2008
  • Place of Presentation: Kanazawa
  • Year and Date: 2008-01-24
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Reconstitution of hepatic tissue architectures from mouse liver cells using simulated microgravity culture system (2007)
  • Author(s): Ueno, Y., Okamur, A., Zheng, Y.W., Tanaka, J., Tateisi, T., Tahiguchi, H
  • Organizer: The China-Japan Mini-Symposium 2007 on Nanotechnology and Tissue Enginieering
  • Place of Presentation: Japan
  • Year and Date: 2007-12-08
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Excessive self-renewal of hepaticatem cells derives cancer initiation (2007)
  • Author(s): Taniguchi H
  • Organizer: 2007 Soon Chun hyang Stem Cell Symposium
  • Place of Presentation: Seoul
  • Year and Date: 2007-09-15
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Excessive self-renewal of hepatic stem/progenitor cells derives cancer initiation (2007)
  • Author(s): Taniguchi, H
  • Organizer: 2007Soon Chun hyang Stem Cell Symposium
  • Place of Presentation: Seoul
  • Year and Date: 2007-09-15
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Excessive self-renewal of hepatic stem cells derives cancer initiation (2007)
  • Author(s): Taniguchi H
  • Organizer: 2007 Soon Chun hyang Stem Cell Symposium
  • Place of Presentation: Seoul
  • Year and Date: 2007-09-15
• [Presentation] In vitro propagation of multipotent intestinal stem cells (2007)
  • Author(s): Suzuki, A., Sekiya, S., Taniguchi, H
  • Organizer: 12th US-Japan G1 & Liver Meeting in 21st Century
  • Place of Presentation: Kyoto
  • Year and Date: 2007-06-22
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 膵島創出へ向けた細胞操作技術の開発 (2007)
  • Author(s): 谷口 英樹、仲野 晶、岡村 愛、田中 順三
  • Organizer: 第34回日本臓器保存生物医学会学術集会
  • Place of Presentation: 札幌
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Excessive self-renewal of hepaticatem/progenitor cells drives cancer initiation (2007)
  • Author(s): Taniguchi H
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] ヒト研究用モデル細胞としての幹細胞クローン (2007)
  • Author(s): 谷口 英樹
  • Organizer: 第25回日本ヒト細胞学会学術集会
  • Place of Presentation: 東京
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 膵幹細胞の可視化による非対称分裂の解析 (2007)
  • Author(s): 谷口 英樹
  • Organizer: 第17回日本サイトメトリー学会学術集会
  • Place of Presentation: 東京
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 組織幹細胞と"がん幹細胞" (2007)
  • Author(s): 谷口 英樹
  • Organizer: 第19回日本肝胆膵外科学会・学術集会
  • Place of Presentation: 横浜
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Excessive self-renewal of hepatic: stem/progenitor cells drives cancer initiation (2007)
  • Author(s): Taniguchi H
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
• [Presentation] Clonal Identification of Mulltipotent Stem/Progenitor Cells in the Developing Liver and Pancreas Using flowcytometric Cell Sorting. (2006)
  • Author(s): Taniguchi H
  • Organizer: The China-Japan Mini-Symposium 2005 on Regenerative Medecine
  • Place of Presentation: Shanghai
  • Year and Date: 2006-10-21
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 外科領域への幹細胞生物学のインパクト (2006)
  • Author(s): 谷口 英樹
  • Organizer: 第33回日本臓器保存生物医学会総会
  • Place of Presentation: 東京
  • Year and Date: 2006-09-23
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 組織幹細胞を用いた眼科プロセスの再構成 (2006)
  • Author(s): 谷口 英樹
  • Organizer: 第95回日本病理学会総会
  • Place of Presentation: 東京
  • Year and Date: 2006-08-30
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 肝幹細胞を対象とした腫瘍化プロセスの再構成 (2006)
  • Author(s): 谷口 英樹
  • Organizer: 第13回肝細胞研究会
  • Place of Presentation: 旭川
  • Year and Date: 2006-06-30
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 固形臓器における幹細胞システムの解明 (2006)
  • Author(s): 谷口 英樹
  • Organizer: 第83回日本生理学会大会
  • Place of Presentation: 前橋
  • Year and Date: 2006-03-28
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 固形臓器における組織幹細胞の特性解析 (2006)
  • Author(s): 谷口 英樹
  • Organizer: 第2回宮崎サイエンスキャンプ
  • Place of Presentation: 宮崎
  • Year and Date: 2006-02-24
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Clonal identification of multipotent stem/progenitor cells in the liver and pancreas using flowcytometric cell sorting (2006)
  • Author(s): Taniguchi, H
  • Organizer: The 5th JBS Symposium
  • Place of Presentation: Karuizawa
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Clonal Identification of Mulltipotent Stem/Progenitor Cells in the Develping Liver and Pancreas Using flowcytometric Cell Sorting (2005)
  • Author(s): Taniguchi, H
  • Organizer: The China-Japan Mini-Symposium 2005 on Regenerative Medecine
  • Place of Presentation: Shanghai. China
  • Year and Date: 2005-10-21
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Clonal Identification of Mulltipotent Stem/Progenitor Cells in the Liver and Pancreas Using Flowcytometric Cell sorting. (2005)
  • Author(s): Taniguchi H
  • Organizer: Preparing for New Centennial Medicine
  • Place of Presentation: Seoul
  • Year and Date: 2005-09-03
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 再生医療に向けた固形臓器における幹細胞研究の現状 (2005)
  • Author(s): 谷口 英樹
  • Organizer: 第21回日本DDS学会
  • Place of Presentation: 長崎
  • Year and Date: 2005-07-22
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Clonal Identification of Mulltipotent Stem/Progenitor Cells in The Developing Liver and Pancreas Using Flowcytometric Cell Sorting (2005)
  • Author(s): Taniguchi H
  • Organizer: International Symposium on Soft-Nanotechnology 2005
  • Place of Presentation: Sapporo
  • Year and Date: 2005-06-20
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Clonal identification of mulltipotent stem/progenitor cells in the developing liver and pancreas using flowcytometric cell sorting (2005)
  • Author(s): 谷口 英樹
  • Organizer: 2005 Trilateral Stem Cell Meeting-Japan, France, Pittsburgh
  • Place of Presentation: Kobe
  • Year and Date: 2005-05-12
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Clonal identification of mulltipotent stem/progenitor cells in the liver and pancreas using flowcytometric cell sorting. (2005)
  • Author(s): Taniguhi H
  • Organizer: The 5th JBS Symposium
  • Place of Presentation: Karuizawa
  • Year and Date: 2005-05-01
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Clonal Identification of Multipotent Stem/Progenitor Cells in the Liver and Pancreas Using Flowcytometric Cell sorting (2005)
  • Author(s): Taniguchi, H
  • Organizer: Preparing for New Centennial Medicine Sympsium
  • Place of Presentation: Seoul
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Clonal Identification of Mulltipotent Stem/Progenitor Soft-Nanotechnology 2005, Cells in The Developing Liver and Pancreas Using Flowcytometric Cell Sorting (2005)
  • Author(s): Taniguchi, H
  • Organizer: International Symposium on
  • Place of Presentation: Sapporo, Japan
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Clonal identification of mullipotent stem/progenitor cell in the developing liver and pancreas using flowcytometric cell sorting (2005)
  • Author(s): Taniguchi, H
  • Organizer: 2005 Trilateral Stem Cell Meeting-Japan,France, Pittsburgh-
  • Place of Presentation: Kobe
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Hedgehog signaling is repuired for maintenance of neonatal pancreaticstem/progenitor cells. (2004)
  • Author(s): Zhao W.Ishigawa M.Hirose T.Chiba T.Taniguchi H
  • Organizer: 第28回日本分子生物学会年会
  • Place of Presentation: 福岡
  • Year and Date: 2004-12-07
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 再生医孫工学の技術 (2007)
  • Author(s): 谷口 英樹(分担執筆)
  • Total Pages: 251
  • Publisher: シーエムシー出版
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 再生医療工学の技術 (2007)
  • Author(s): 谷口英樹(分担執筆)
  • Total Pages: 251
  • Publisher: シーエムシー出版
• [Book] 胆管上皮細胞の癌化機構; 日本臨床増刊号 膵癌・胆道癌の診断と治療(共同執筆) (2006)
  • Author(s): 谷口 英樹
  • Total Pages: 6
  • Publisher: 日本臨床社
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 肝臓再生を目指して; 人体再生 幹細胞がひらく未来の医療 別冊日経サイエンス(共同執筆) (2006)
  • Author(s): 谷口 英樹
  • Total Pages: 4
  • Publisher: 日経サイエンス社
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 糖尿病を再生医療で治す; 人体再生 幹細胞がひらく未来の医療 別冊日経サイエンス(共同執筆) (2006)
  • Author(s): 谷口 英樹
  • Total Pages: 6
  • Publisher: 日経サイエンス社
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 肝臓における幹細胞と癌の接点; 肝疾患Review2006-2007(共同執筆) (2006)
  • Author(s): 谷口 英樹
  • Total Pages: 6
  • Publisher: 日本メディカルセンター
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 胆管上皮細胞の癌化機構;日本臨床増刊号膵癌・胆道癌の診断と治療(共同執筆) (2006)
  • Author(s): 谷口英樹
  • Total Pages: 6
  • Publisher: 日本臨床社
• [Book] 肝臓再生を目指して;人体再生 幹細胞がひらく未来の医療 別冊日経サイエンス(共同執筆) (2006)
  • Author(s): 谷口英樹
  • Total Pages: 4
  • Publisher: 日経サイエンス社
• [Book] 糖尿病学2005(膵臓における組織幹細胞の分離と再生治療への展望) (2005)
  • Author(s): 谷口 英樹
  • Total Pages: 8
  • Publisher: 診断と治療社
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 糖尿病学2005(膵臓における組織幹細胞の分離と再生治療への展望) (2005)
  • Author(s): 谷口英樹
  • Total Pages: 8
  • Publisher: 診断と治療社