Reconstitution of human haematopoiesis in non-obese diabetic/severe combined immunodeficient mice by clonal cells expanded from single CD34+CD38-cells expressing Flk2/Flt3

Yasuhiro Ebihara, Mika Wada, Takahiro Ueda, Mingjiang Xu, Atsushi Manabe, Ryuhei Tanaka, Mamoru Ito, Hideo Mugishima, Shigetaka Asano, Tatsutoshi Nakahata, Kohichiro Tsuji

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24 Scopus citations

Abstract

In the present study, we examined the expression of Flk2/Flt3, a tyrosine kinase receptor, on human cord blood CD34+ haematopoietic progenitor/stem cells. In flow cytometric analysis, Flk2/Flt3 was expressed on 80% of CD34+ cells and their immature subpopulations, CD34+CD33- and CD34+CD38- cells. Methycellulose clonal culture of sorted CD34+Flk2/Flt3+ and CD34+Flk2/Flt3- cells showed that most of myelocytic progenitors expressed Flk2/Flt3, but erythroid and haematopoietic multipotential progenitors were shared by both fractions. When 1 × 104 lineage marker-negative (Lin-)CD34+Flk2/Flt3- cells were transplanted into non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice, none of the recipients possessed human CD45+ cells in bone marrow 11-12 weeks after the transplantation. In contrast, all recipients transplanted with 1 × 104 Lin-CD34+Flk2/Flt3+ cells showed successful engraftment. Furthermore, clonal cells expanded from single Lin-CD34+CD38-Flk2/Flt3+ cells in the culture with Flk2/Flt3 ligand, stem cell factor, thrombopoietin, and a complex of interleukin 6/soluble interleukin 6 receptor were individually transplanted into NOD/SCID mice. At 20 to 21 weeks after the transplantation, three out of 10 clones harvested at d 7 of culture, and three out of six clones at d 14 could reconstitute human haematopoiesis in recipient marrow. These results demonstrated that Flk2/Flt3 was expressed on a wide variety of human haematopoietic cells including long-term-repopulating haematopoietic stem cells.

Original languageEnglish (US)
Pages (from-to)525-534
Number of pages10
JournalBritish Journal of Haematology
Volume119
Issue number2
DOIs
StatePublished - 2002
Externally publishedYes

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Keywords

  • CD34
  • Cord blood
  • Flk2/Flt3
  • Long-term-repopulating haematopoietic stem cells (LTR-HSC)
  • NOD/SCID mice

ASJC Scopus subject areas

  • Hematology

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