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 Blood, 15 August 2004, Vol. 104, No. 4, pp. 1002-1009.
Prepublished online as a Blood First Edition Paper on April 22, 2004; DOI 10.1182/blood-2003-09-3347.

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HEMATOPOIESIS

Indian hedgehog gene transfer augments hematopoietic support of human stromal cells including NOD/SCID–ß2m–/– repopulating cells.

Masayoshi Kobune, Yoshinori Ito, Yutaka Kawano, Katsunori Sasaki, Hiroaki Uchida, Kiminori Nakamura, Hironari Dehari, Hiroki Chiba, Rishu Takimoto, Takuya Matsunaga, Takeshi Terui, Junji Kato, Yoshiro Niitsu, and Hirofumi Hamada

From the Fourth Department of Internal Medicine, the Department of Molecular Medicine, and the Division of Gene Therapy, Sapporo Medical University School of Medicine, Sapporo, Japan; and the Department of Surgery and Bioengineering, Institute of Medical Science, University of Tokyo, Tokyo, Japan.

Hematopoietic stem cells (HSCs) are a subset of bone marrow cells that are capable of self-renewal and of giving rise to all types of blood cells. However, the mechanisms involved in controlling the number and abilities of HSCs remain largely unknown. The Indian hedgehog (Ihh) signal has an essential role in inducing hematopoietic tissue during embryogenesis. We investigated the roles of the Ihh in coculture with CD34+ cells and human stromal cells. Ihh mRNA was expressed in primary and telomerized human (hTERT) stromal cells, and its receptor molecules were detected in CD34+ cells. Ihh gene transfer into hTERT stromal cells enhanced their hematopoietic supporting potential, which was elevated compared with control stromal cells, as indicated by the colony-forming units in culture (CFU-Cs) (26-fold ± 2-fold versus 59-fold ± 3-fold of the initial cell number; mixed colony-forming units [CFU-Mix's], 63-fold ± 37-fold versus 349-fold ± 116-fold). Engraftments of nonobese diabetic/severe combined immunodeficiency–ß2m–/– (NOD/SCID–ß2m–/–) repopulating cells (RCs) expanded on Ihh stromal cells were significantly higher compared with control coculture results, and engraftment was neutralized by addition of an antihedgehog antibody. Limiting dilution analysis indicated that NOD/SCID–ß2m–/– RCs proliferated efficiently on Ihh stromal cells, compared with control stromal cells. These results indicate that Ihh gene transfer could enhance the primitive hematopoietic support ability of human stromal cells.


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