The Stem Cell Core will support all projects by providing expertise in the purification and characterization of primitive hematopoietic cell populations as well as the tracking of genetically modified cells in bone marrow and peripheral blood following reconstitution. Expertise has been established in the preparation of primitive hematopoietie cell populations from three species: humans, rhesus macaques and mice. Human peripheral blood cells obtained from normal volunteers following administration of G-CSF are purified using the Clinimacs and/or the Aria cell sorter. The shared resource is equipped with Miltenyi MACS system as well as a state-of-the-art Becton-Dickinson Aria cell sorter. The Clinimacs is also used to recover CD34 + cells from peripheral blood or bone marrow of rhesus macaques following administration of G-CSF and SCF. Methodologies depending on depletion of cells expressing lineage markers are being developed using the AutoMACS device with enrichment in primitive cells monitored by quantitating the side population (SP) cells defined by dye efflux. Two methodologies are available within the Core for preparation of cell populations highly enriched in murine repopulating stem cells. The first involves the use of dye efflux to isolate the SP population. The expertise in purifying functional SP cells will be adapted for use in the rhesus model. The methodology is sufficiently developed to allow reconstitution of mice with as few as 250 transplanted SP calls. The second strategy involves selection of lineage -ckit+, Sca1+ cell by FACS. Overall, the Core provides essential expertise, instrumentation and experience in complex methodologies necessary to purify, eharacterize and analyze primitive hematopoietic cell populations.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL053749-14
Application #
7487356
Study Section
Heart, Lung, and Blood Initial Review Group (HLBP)
Project Start
Project End
Budget Start
2007-09-01
Budget End
2008-08-31
Support Year
14
Fiscal Year
2007
Total Cost
$129,910
Indirect Cost
Name
St. Jude Children's Research Hospital
Department
Type
DUNS #
067717892
City
Memphis
State
TN
Country
United States
Zip Code
38105
Zhao, Hui Fen; Abraham, Allistair; Kim, Yoon-Sang et al. (2017) Lentiviral Transfer of ?-Globin with Fusion Gene NUP98-HOXA10HD Expands Hematopoietic Stem Cells and Ameliorates Murine ?-Thalassemia. Mol Ther 25:593-605
De Ravin, Suk See; Wu, Xiaolin; Moir, Susan et al. (2016) Lentiviral hematopoietic stem cell gene therapy for X-linked severe combined immunodeficiency. Sci Transl Med 8:335ra57
Abraham, Allistair; Kim, Yoon-Sang; Zhao, Huifen et al. (2016) Increased Engraftment of Human Short Term Repopulating Hematopoietic Cells in NOD/SCID/IL2r?null Mice by Lentiviral Expression of NUP98-HOXA10HD. PLoS One 11:e0147059
Wielgosz, Matthew M; Kim, Yoon-Sang; Carney, Gael G et al. (2015) Generation of a lentiviral vector producer cell clone for human Wiskott-Aldrich syndrome gene therapy. Mol Ther Methods Clin Dev 2:14063
Pestina, Tamara I; Hargrove, Phillip W; Zhao, Huifen et al. (2015) Amelioration of murine sickle cell disease by nonablative conditioning and ?-globin gene-corrected bone marrow cells. Mol Ther Methods Clin Dev 2:15045
Zhou, Sheng; Bonner, Melissa A; Wang, Yong-Dong et al. (2015) Quantitative shearing linear amplification polymerase chain reaction: an improved method for quantifying lentiviral vector insertion sites in transplanted hematopoietic cell systems. Hum Gene Ther Methods 26:4-12
Urbinati, Fabrizia; Hargrove, Phillip W; Geiger, Sabine et al. (2015) Potentially therapeutic levels of anti-sickling globin gene expression following lentivirus-mediated gene transfer in sickle cell disease bone marrow CD34+ cells. Exp Hematol 43:346-351
Yu, Hui; Neale, Geoffrey; Zhang, Hui et al. (2014) Downregulation of Prdm16 mRNA is a specific antileukemic mechanism during HOXB4-mediated HSC expansion in vivo. Blood 124:1737-47
Treanor, Louise M; Zhou, Sheng; Janke, Laura et al. (2014) Interleukin-7 receptor mutants initiate early T cell precursor leukemia in murine thymocyte progenitors with multipotent potential. J Exp Med 211:701-13
Griffith, Linda M; Cowan, Morton J; Notarangelo, Luigi D et al. (2014) Primary Immune Deficiency Treatment Consortium (PIDTC) report. J Allergy Clin Immunol 133:335-47

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