Development of the lymphocyte in vitro amplification method using HHIP KO stromal cells.

• Project/Area Number: 24591427
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Hematology
• Research Institution: Sapporo Medical University
• Principal Investigator: IYAMA Satoshi 札幌医科大学, 医学部, 助教 (50398319)
• Co-Investigator(Kenkyū-buntansha): KATO Junji 札幌医科大学, 医学部, 教授 (20244345) ; KOBUNE Masayoshi 札幌医科大学, 医学部, 准教授 (90336389) ; KAMIHARA Yusuke 札幌医科大学, 医学部, 研究員 (10624421)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: リンパ球増幅 / 造血幹細胞

Outline of Final Research Achievements

We have previously shown that it was difficult to evaluate 5 weeks CA-forming cells (CAFC) which is a surrogate indicator of hematopoietic stem cells because human stromal cells could not survive more than 4 weeks. In this study, we optimized surum free media, combination of cytokines and chemical agents such as stem cell factor, thrombopoietin, flt3 ligand with or without delta like protein 4 (DLL4) and GSK inhibitor. By using this media, bone marrow CD34+ cells could be cocultured for 8 weeks without disruption of the human stromal layer. Five week CAFC could be observed in all condition of cytokine combination with or without chemical inhibitor. However, because cytoplasmic appearance of cells in some CAs is quite irregular without DLL4 or GSK inhibitor, we conducted immunohistochemical staining. We found that CD34+CAFCs were exclusively observed in the presence of DLL4 or GSK inhibitor. Moreover, NOD/SCID repopulating cells were detected in the coculture containing CD34+CAFCs.

Research Products

• [Journal Article] Narrowband ultraviolet B phototherapy ameliorates acute graft-versus-host disease by a mechanism involving in vivo expansion of CD4+CD25+Foxp3+ regulatory T cells． (2014)
  • Author(s): Iyama S，Murase K，Sato T，Hashimoto A，Tatekoshi A，Horiguchi H，Kamihara Y，Ono K，Kikuchi S，Takada K，Kawano Y，Hayashi T，Miyanishi K，Sato Y，Takimoto R，Kobune M，Mori S，Kato J，Yamashita T，Kato J．
  • Journal Title: Int J Hematol Volume: 99 Issue: 4 Pages: 471-476
  • DOI: 10.1007/s12185-014-1530-1
  • Peer Reviewed
• [Journal Article] Efficacy of Enteral Supplementation Enriched with Glutamine, Fiber, and Oligosaccharide on Mucosal Injury following Hematopoietic Stem Cell Transplantation (2014)
  • Author(s): Iyama S, Sato T, Tatsumi H, Hashimoto A, Tatekoshi A, Kamihara Y, Horiguchi H, Ibata S, Ono K, Murase K, Takada K, Sato Y, Hayashi T, Miyanishi K, Akizuki E, Nobuoka T, Mizuguchi T, Takimoto R, Kobune M, Hirata K, Kato J.
  • Journal Title: Case Rep Oncol Volume: 7 Issue: 3 Pages: 692-699
  • DOI: 10.1159/000368714
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Exosomes Derived from AML/MDS Cells Is Involved in Stromal Dysfunction and Bone Marrow Failure. (2014)
  • Author(s): Horiguchi H, Kobune M, Kikuchi S, Jomen W, Murase K, Ibata S, Iyama S, Sato T, Kamihara Y, Miyanishi K, Sato Y, Hayashi T, Takimoto R, Kato J.
  • Organizer: 56th ASH Annual Meeting and Exposition
  • Place of Presentation: San Francisco, U.S.A
  • Year and Date: 2014-12-06 – 2014-12-09