Establishment of a xeno-free primary culture method for human colonic epithelial stem cells

• Project/Area Number: 25860524
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Gastroenterology
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: MIZUTANI Tomohiro 東京医科歯科大学, 医歯(薬)学総合研究科, 特任助教 (80632588)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 腸管上皮培養 / 低分子化合物 / GMP規格 / 再生医療 / 下部消化管学（小腸、大腸） / 人工ペプチド

Outline of Final Research Achievements

We have developed a novel culture method for human colonic epithelial cells that allows long-term maintenance and efficient expansion of stem cells in vitro from tiny biopsy specimens. The factors used in this method were successfully replaced to GMP (Good Manufacturing Practice)-grade recombinant proteins designed to use for clinical regenerative medicine. We also refined the transplantation method for colonic epithelial cells onto colon and developed a new method for epithelial cell transplantation. With this method, we found that cultured small intestinal cells could be heterotopically engrafted onto the colon, which means that the method would be a good model for cultured human colonic cell transplantation.
These results would be of significant help to build a basis for regenerative medicine using cultured human colonic epithelial stem cells.

Research Products

• [Journal Article] Small intestinal stem cell identity is maintained with functional Paneth cells in heterotopically grafted epithelium onto the colon (2014)
  • Author(s): Fukuda M, Mizutani T, Mochizuki W, Matsumoto T, Nozaki K, Sakamaki Y, Ichinose S, Okada Y, Tanaka T, Watanabe M, Nakamura T
  • Journal Title: Genes & Development Volume: 28(16) Issue: 16 Pages: 1752-1757
  • DOI: 10.1101/gad.245233.114
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Hes1 promotes the IL-22-mediated antimicrobial response by enhancing STAT3-dependent transcription in human intestinal epithelial cells (2014)
  • Author(s): Murano T, Okamoto R, Ito G, Nakata T, Hibiya S, Shimizu H, Fujii S, Kano Y, Mizutani T, Yui S, Akiyama-Morio J, Nemoto Y, Tsuchiya K, Nakamura T, Watanabe M
  • Journal Title: Biochem Biophys Res Commun Volume: 443 Issue: 3 Pages: 840-846
  • DOI: 10.1016/j.bbrc.2013.12.061
  • Peer Reviewed / Open Access
• [Journal Article] Fluorescent labelling of intestinal epithelial cells reveals independent long-lived intestinal stem cells in a crypt. (2014)
  • Author(s): N. Horita, K. Tsuchiya, R. Hayashi, K. Fukushima, S. Hibiya, M. Fukuda, Y. Kano, T. Mizutani, Y. Nemoto, S. Yui, R. Okamoto, T. Nakamura, M. Watanabe
  • Journal Title: Biochem Biophys Res Commun Volume: 454 Issue: 4 Pages: 493-499
  • DOI: 10.1016/j.bbrc.2014.10.091
  • Peer Reviewed / Open Access
• [Journal Article] Distinct expression patterns of Notch ligands, Dll1 and Dll4, in normal and inflamed mice intestine. (2014)
  • Author(s): Shimizu H, Okamoto R, Ito G, Nakata T, Suzuki K, Murano T, Mizutani T, Tsuchiya K, Nakamura T, Hozumi K, Watanabe M.
  • Journal Title: PeerJ Volume: 2 Pages: e370-e370
  • DOI: 10.7717/peerj.370
  • Peer Reviewed / Open Access
• [Journal Article] Lineage-specific expression of Bestrophin-2 and Bestrophin-4 in human intestinal epithelial cells. (2013)
  • Author(s): Ito G, Okamoto R, Nakamura T, Watanabe M et al
  • Journal Title: PloS One. Volume: 8 Issue: 11 Pages: e79693-e79693
  • DOI: 10.1371/journal.pone.0079693
  • Peer Reviewed
• [Presentation] Successful Engraftment of Cultured Small Intestinal Epithelial Stem Cells onto Damaged Colonic Mucosa by Heterotopic Transplantation. (2014)
  • Author(s): Fukuda M, Mizutani T, Mochizuki W, Matsumoto T, Nozaki K, Ichinose S, Watanabe M, Nakamura T.
  • Organizer: ISSCR 12th Annual Meeting
  • Place of Presentation: Vancouver,Canada
  • Year and Date: 2014-06-19
• [Presentation] Successful Engraftment of Cultured Small Intestinal Epithelial Stem Cells onto Damaged Colonic Mucosa by Heterotopic Transplantation. (2014)
  • Author(s): Mizutani T, Fukuda M, Mochizuki W, Matsumoto T, Nozaki K, Ichinose S, Watanabe M, Nakamura T.
  • Organizer: DDW2014
  • Place of Presentation: Chicago,USA
  • Year and Date: 2014-05-03