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

2014 Fiscal Year Final Research Report

• 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
• 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.

Free Research Field

血液学