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Conditions that enable human hematopoietic stem cell engraftment in all NOD-SCID mice
A M Rice1, J A Wood, C G Milross
1Stem Cell Biology Program, Children's Cancer Institute Australia for Medical Research, Radiation Oncology, Prince of Wales Hospital, Randwick, NSW.
Transplantation
|February 7, 2001
Summary
This study defines optimal conditions for human hematopoietic stem cell engraftment in NOD-SCID mice. Researchers identified specific irradiation levels and injection methods for reliable quantification of human stem cells.
Area of Science:
- Hematology
- Immunology
- Stem Cell Biology
Background:
- Evaluating long-term repopulation potential of human hematopoietic stem cells (HSCs) is crucial.
- Existing models for quantifying human HSCs are limited.
- The NOD-SCID mouse model shows promise for human HSC quantification, but engraftment conditions require definition.
Purpose of the Study:
- To define variables enabling consistent human stem cell engraftment in NOD-SCID mice.
- To establish accurate techniques for quantifying human stem cell engraftment.
Main Methods:
- NOD-SCID mice were irradiated (250-400 cGy) and received cord blood (CB) cells (intravenous or intraperitoneal).
- Engraftment was assessed in bone marrow, spleen, and thymus 6 weeks post-transplantation.
- Techniques included four-color flow cytometry, semi-quantitative PCR, and progenitor/stem cell assays.
Main Results:
- Intravenous injection of CB mononuclear or CD34+ cells with 250 or 325 cGy irradiation is necessary for multilineage human engraftment.
- Flow cytometry and PCR accurately detect 0.1% human cells.
- Functional assays provide insights into engrafted cell capacity.
Conclusions:
- The NOD-SCID mouse model, with defined parameters, reliably supports human stem cell engraftment.
- This model and assessment techniques facilitate analysis of cytokine effects on cord blood stem cell repopulating capacity.