We propose to identify and purify human hematopoietic stem cells with long-term engraftment capability. Initially, the major effort will be made to critically examine the dogma that the human hematopoietic stem cells are CD34+. This concept was seriously challenged by recent reports that murine hematopoietic stem cells with long-term engraftment capability are CD34- whereas the CD34+ cells provide short-term to mid- term engraftment in lethally irradiated mice. Using sheep/human xenograft model, we have obtained preliminary information that there are cells capable of long-term engraftment in the CD34-human marrow cell population. It is possible that human stem cells are primarily in the CD34- cell population. It is also possible that the human stem cells are in both CD34+ and CD34-cell populations. Alternatively, there may be a species difference in surface markers between human and murine hematopoiesis. Finally, this apparent controversy may be explainable on the basis of epitope differences of the anti-CD34 antibodies. We will clarify the status of CD34 expression by long-term and short- term engrafting cells and develop alternative methods for purification of human stem cells. We will also correlate the engraftment capabilities of the cell populations with the incidences of long-term culture-initiating cells (LTC-IC) and blast cell colony progenitors. Two types of xenograft assays will be used for quantitation of human engrafting cells-sheep/ human chimera and NOD/SCID mice. The sheep/human xenograft assay is particularly suited for the measurement of human long-term engrafting cells. NOD/SCID mouse model is more practical for limiting dilution studies. We already have exciting preliminary data regarding the use of myeloablated NOD/SCID newborn mice for quantitation of human engrafting cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK054197-04
Application #
6381174
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Badman, David G
Project Start
1998-08-14
Project End
2003-07-31
Budget Start
2001-08-01
Budget End
2003-07-31
Support Year
4
Fiscal Year
2001
Total Cost
$316,951
Indirect Cost
Name
Medical University of South Carolina
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
183710748
City
Charleston
State
SC
Country
United States
Zip Code
29425
Abe, Takanori; Masuya, Masahiro; Ogawa, Makio (2010) An efficient method for single hematopoietic stem cell engraftment in mice based on cell-cycle dormancy of hematopoietic stem cells. Exp Hematol 38:603-8
Minamiguchi, Hitoshi; Ishikawa, Fumihiko; Fleming, Paul A et al. (2008) Transplanted human cord blood cells generate amylase-producing pancreatic acinar cells in engrafted mice. Pancreas 36:e30-5
Masuya, Masahiro; Moussa, Omar; Abe, Takanori et al. (2005) Dysregulation of granulocyte, erythrocyte, and NK cell lineages in Fli-1 gene-targeted mice. Blood 105:95-102
Minamiguchi, Hitoshi; Wingard, John R; Laver, Joseph H et al. (2005) An assay for human hematopoietic stem cells based on transplantation into nonobese diabetic recombination activating gene-null perforin-null mice. Biol Blood Marrow Transplant 11:487-94
Abe, Takanori; Fleming, Paul A; Masuya, Masahiro et al. (2005) Granulocyte/macrophage origin of glomerular mesangial cells. Int J Hematol 82:115-8
Hess, David C; Abe, Takanori; Hill, William D et al. (2004) Hematopoietic origin of microglial and perivascular cells in brain. Exp Neurol 186:134-44
Higuchi, Y; Zeng, H; Ogawa, M (2003) CD38 expression by hematopoietic stem cells of newborn and juvenile mice. Leukemia 17:171-4
Zanjani, Esmail D; Almeida-Porada, Graca; Livingston, Anne G et al. (2003) Reversible expression of CD34 by adult human bone marrow long-term engrafting hematopoietic stem cells. Exp Hematol 31:406-12
Ishikawa, Fumihiko; Drake, Christopher J; Yang, Su et al. (2003) Transplanted human cord blood cells give rise to hepatocytes in engrafted mice. Ann N Y Acad Sci 996:174-85
Ishikawa, F; Livingston, A G; Minamiguchi, H et al. (2003) Human cord blood long-term engrafting cells are CD34+ CD38-. Leukemia 17:960-4

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