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Hematopoietic Repopulating Ability of Cord Blood CD34⁺ Cells in NOD/Shi-scid Mice

^a Department of Clinical Oncology and
^b Department of Molecular Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan;
^c Central Institute for Experimental Animals, Kawasaki, Japan;
^d Department of Pediatrics, Kyoto University, Kyoto, Japan

Key Words. Hematopoietic repopulating ability • Cord blood • CD34⁺ cells • NOD/Shi-scid mice • Hematopoietic stem cells • Hematopoietic stem cell transplantation

Correspondence: Kohichiro Tsuji, M.D., Department of Clinical Oncology, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan. Telephone: +81-3-5449-5397; Fax: +81-3-5449-5428; e-mail: tsujik{at}ims.u-tokyo.ac.jp

Although umbilical cord blood (CB) is increasingly being used as an alternative to bone marrow (BM) as a source of transplantable hematopoietic stem cells (HSC), information on the hematopoietic repopulating ability of CB HSC is still limited. We recently established a xenotransplantation system in NOD/Shi-scid mice to evaluate human stem cell activity. In the present study, we transplanted 5 to 10 x 10⁴ CB CD34⁺ cells into six NOD/Shi-scid mice treated with anti-asialo GM1 antiserum to investigate the hematopoietic repopulating ability of CB. The BM of all recipients contained human CD45⁺ cells 10 to 12 weeks after the transplantation (43.8 ± 17.7%). Clonal culture of the recipient BM cells revealed the formation of various types of human hematopoietic colonies, including myelocytic, erythroid, megakaryocytic, and multilineage colonies, indicating that CB HSC can differentiate into hematopoietic progenitors of various lineages. However, the extent of the differentiation and maturation differed with each lineage. CD13⁺/CD14⁺/CD33⁺ myelocytic cells were mainly repopulated in BM and peripheral blood (PB). While CD41⁺ megakaryocytic cells and platelets were present, few glycophorin A⁺CD71⁺ or hemoglobin -containing erythroid cells were detected. CD19⁺ B cells were the most abundantly repopulated in NOD/Shi-scid mice, but their maturational stage differed among the hematopoietic organs. Most of the BM CD19⁺ cells were immature B cells expressing CD10 but not surface immunoglobulin (Ig) M, whereas more mature CD19⁺CD10^– surface IgM⁺ B cells were predominantly present in spleen and PB. CD3⁺ T cells were not detected even in the recipient thymus. The transplantation to the NOD/Shi-scid mouse may provide a useful tool for evaluating the repopulating ability of transplantable human HSC.

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