title: Conditions that enable human hematopoietic stem cell engraftment in all NOD-SCID mice.
abstract: BACKGROUND
  Transplantation of human hematopoietic stem cells is the only true test of their long-term repopulation potential. Models are readily available to evaluate murine hematopoietic stem cells, but few exist that allow reliable quantification of human stem cells. The non-obese diabetic-severe combined immunodeficient (NOD-SCID) mouse model enables quantification of human hematopoietic stem cells, but the conditions that permit human engraftment in all animals have yet to be defined. The aims of the project were, therefore, to describe the variables that allow human engraftment in the NOD-SCID mouse model and the techniques that accurately quantify this engraftment.
  
  
  METHODS
  NOD-SCID mice that had or had not received 250, 325, or 400 cGy irradiation received cord blood (CB) mononuclear or CD34+ cells i.v. or i.p. Mice were killed 6 weeks after transplantation, and the bone marrow, spleen, and thymus were harvested. Four-color flow cytometric analysis, semi-quantitative PCR, myeloid and erythroid progenitor, and stem cell assays were used to monitor human engraftment.
  
  
  RESULTS
  A 250 or 325 cGy and i.v. injection of CB mononuclear or CD34+ cells is required to detect multilineage human engraftment in the bone marrow, spleen, or thymus of NOD-SCID mice. Four-color flow cytometric analysis and semi-quantitative PCR enable accurate detection of 0.1% human cells. Progenitor and stem cell assays provide functional information about the engrafted cells.
  
  
  CONCLUSIONS
  Successful development of the NOD-SCID mouse model and techniques to assess human engraftment now allow it to be used reliably to analyze the effects of short-term cytokine exposure on the long-term repopulating capacity of CB stem cells.
authors.affiliation: Stem Cell Biology Program, Children's Cancer Institute Australia for Medical Research, Radiation Oncology, Prince of Wales Hospital, Randwick, NSW.
authors.forename: A M
authors.initials: AM
authors.lastname: Rice
authors.forename: J A
authors.initials: JA
authors.lastname: Wood
authors.forename: C G
authors.initials: CG
authors.lastname: Milross
authors.forename: C J
authors.initials: CJ
authors.lastname: Collins
authors.forename: N F
authors.initials: NF
authors.lastname: McCarthy
authors.forename: M R
authors.initials: MR
authors.lastname: Vowels
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis; D004307:Dose-Response Relationship, Radiation; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000276:immunology; D006801:Humans; D006824:Hybridization, Genetic; D007267:Injections; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D011827:Radiation; D013997:Time Factors; D014183:Transplantation, Heterologous
chemical_list: D018952:Antigens, CD34