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- Title
Identification of Long-Term Repopulating Potential of Human Cord Blood-Derived CD34¯flt3¯ Severe Combined Immunodeficiency-Repopulating Cells by Intra-Bone Marrow Injection.
- Authors
Kimura, Takafumi; Asada, Rumiko; Jianfeng Wang; Kimura, Takashi; Morioka, Miho; Matsui, Kazuo; Kobayashi, Katsuya; Henmi, Kae; Imai, Shiro; Kita, Masakazu; Tsuji, Takashi; Sasaki, Yutaka; Ikehara, Susumu; Sonoda, Yoshiaki
- Abstract
Recently, we have identified human cord blood (CB)-derived CD34-negative (CD34-) severe combined immunodeficiency (SCID)-repopulating cells (SRCs) using the intra-bone marrow injection (IBMI) method (Blood 2003; 101:2924). In contrast to murine CD34- Kit+Sca-1+Lineage- (KSL) cells, human CB-derived Lin-CD34- cells did not express detectable levels of c-kit by flow cytometry. In this study, we have investigated the function of flt3 in our identified human CB-derived CD34- SRCs. Both CD34+flt3+/- cells showed SRC activity. In the CD34- cell fraction, only CD34-flt3- cells showed distinct SRC activity by IBMI. Although CD34+flt3+ cells showed a rather weak secondary repopulating activity, CD34+flt3- cells repopulated many more secondary recipient mice. However, CD34-flt3- cells repopulated all of the secondary recipients, and the repopulating rate was much higher. Next, we cocultured CD34-flt3- cells with the murine stromal cell line HESS-5. After 1 week, significant numbers of CD34+flt3+/- cells were generated, and they showed distinct SRC activity. These results indicated that CB-derived CD34-flt3- cells produced CD34+flt3- as well as CD34+flt3+ SRCs in vitro. The present study has demonstrated for the first time that CB-derived CD34- SRCs, like murine CD34- KSL cells, do not express flt3. On the basis of these data, we propose that the immunophenotype of very primitive long-term repopulating human hematopoietic stem cells is Lin-CD34-c-kit-flt3-.
- Subjects
CORD blood; BONE marrow; HEMATOPOIETIC stem cells; CELLS; IMMUNODEFICIENCY; FLOW cytometry
- Publication
Stem Cells, 2007, Vol 25, Issue 6, p1348
- ISSN
1066-5099
- Publication type
Article
- DOI
10.1634/stemcells.2006-0727