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Preferential expression of the vasoactive intestinal peptide (VIP) receptor VPAC1 in human cord blood-derived CD34+CD38− cells: possible role of VIP as a growth-promoting factor for hematopoietic stem/progenitor cells

Leukemia volume 18, pages 912–921 (2004)Cite this article

Abstract

Primitive hematopoietic progenitor cells such as severe combined immunodeficiency- repopulating cells and long-term culture-initiating cells are enriched in CD34+CD38− cells derived from various stem cell sources. In this study, to elucidate the features of such primitive cells at the molecular level, we tried to isolate genes that were preferentially expressed in umbilical cord blood (CB)-derived CD34+CD38− cells by subtractive hybridization. The gene for VPAC1 receptor, a receptor for the neuropeptide vasoactive intestinal peptide (VIP), was thereby isolated and it was shown that this gene was expressed in both CD34+CD38− and CD34+CD38+ CB cells and that the expression levels were higher in CD34+CD38− CB cells. Next, we assessed the effects of VIP on the proliferation of CD34+ CB cells using in vitro culture systems. In serum-free single-cell suspension culture, VIP enhanced clonal growth of CD34+ CB cells in synergy with FLT3 ligand (FL), stem cell factor (SCF), and thrombopoietin (TPO). In serum-free clonogenic assays, VIP promoted myeloid (colony-forming unit-granulocyte/macrophage (CFU-GM)) and mixed (CFU-Mix) colony formations. Furthermore, in Dexter-type long-term cultures, VIP increased colony-forming cells at week 5 of culture. These results suggest that VIP functions as a growth-promoting factor of CB-derived hematopoetic progenitor cells.

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Acknowledgements

We are grateful to Kirin Brewery and Immunex Corporation for providing the various growth factors used in this study. We also thank Kumi Honma and Asuka Kishimoto for cryopreservation of cord blood and bone marrow samples and Machiko Mishima for skillful technical assistance. This work was supported by grants-in-aid for scientific research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (No. 11670996).

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Authors and Affiliations

  1. Department of Cancer Immunotherapy, Osaka University Graduate School of Medicine, Osaka, Japan

    M Kawakami, K Ikegame & A Tsuboi

  2. Department of Molecular-Targeting Cancer Prevention, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan

    T Kimura & Y Sonoda

  3. Department of Clinical Laboratory Science, Osaka University Graduate School of Medicine, Osaka, Japan

    Y Kishimoto, Y Oji & H Sugiyama

  4. Department of Medicine, Kinki Central Hospital, Hyogo, Japan

    T Tatekawa

  5. Department of Immunology & Cancer Program, Oklahoma Medical Research Foundation, Oklahoma, USA

    Y Baba

  6. Department of Obstetrics and Gynecology, Suita Municipal Hospital, Osaka, Japan

    T Nishizaki

  7. Department of Obstetrics and Gynecology, Ikeda Municipal Hospital, Osaka, Japan

    N Matsuzaki

  8. Department of Molecular Medicine, Osaka University Graduate School of Medicine, Osaka, Japan

    Y Taniguchi, S Yoshihara, T Shirakata, S Nishida, T Masuda, N Hosen, Y Oka, H Ogawa & I Kawase

  9. Department of Medicine, Osaka Minami National Hospital, Osaka, Japan

    T Soma

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Correspondence to M Kawakami.

Appendix A

Appendix A

Primers and probes for real-time RT-PCR are shown in Table A.1.

Table a1 Primers and probes for real-time RT-PCR
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Kawakami, M., Kimura, T., Kishimoto, Y. et al. Preferential expression of the vasoactive intestinal peptide (VIP) receptor VPAC1 in human cord blood-derived CD34+CD38− cells: possible role of VIP as a growth-promoting factor for hematopoietic stem/progenitor cells. Leukemia 18, 912–921 (2004). https://doi.org/10.1038/sj.leu.2403330

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