Indian hedgehog gene transfer augments hematopoietic support of human stromal cells including NOD/SCID-beta2m-/- repopulating cells.
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-beta2m-/- (NOD/SCID-beta2-/-) 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-beta2m-/- 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.
Kobune
Masayoshi
M
Department of Molecular Medicine, Sapporo Medical University; Chuo-ku S1, W17, Sapporo, 060-8556, Japan. mkobune@ex.me-h.ne.jp
Ito
Yoshinori
Y
Kawano
Yutaka
Y
Sasaki
Katsunori
K
Uchida
Hiroaki
H
Nakamura
Kiminori
K
Dehari
Hironari
H
Chiba
Hiroki
H
Takimoto
Rishu
R
Matsunaga
Takuya
T
Terui
Takeshi
T
Kato
Junji
J
Niitsu
Yoshiro
Y
Hamada
Hirofumi
H
D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D018920:Coculture Techniques; D004268:DNA-Binding Proteins; D005312:Fetal Blood / Q000166:cytology; D006085:Graft Survival; D053823:Hedgehog Proteins; D006410:Hematopoiesis / Q000187:drug effects; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D012333:RNA, Messenger / Q000032:analysis; D017154:Stromal Cells / Q000737:chemistry / Q000166:cytology*; D019098:Telomerase; D015534:Trans-Activators / Q000032:analysis / Q000235:genetics / Q000494:pharmacology*; D014161:Transduction, Genetic
D004268:DNA-Binding Proteins; D053823:Hedgehog Proteins; D012333:RNA, Messenger; D015534:Trans-Activators; D019098:Telomerase