Rhodamine123 reveals heterogeneity within murine Lin-, Sca-1+ hemopoietic stem cells.

Li, C L; Johnson, Gregory R.

doi:10.1084/jem.175.6.1443

Cited by 149 publications

(86 citation statements)

References 10 publications

(17 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These results suggested the existence of a population of stem cells that exhibited only LTRA. These findings contrast other reports employing highly purified stem cell populations that appeared to exhibit good radioprotective capacity [1, [9][10][11][12][13]. The apparent discrepancy between these studies may be explained by the frequency of stem cells in the elutriated cell population reported by Jones.…”

Section: Bone Marrow Repopulation Following Syngeneic Transplantationcontrasting

confidence: 85%

Differential short&hyphen;term and long&hyphen;term repopulating ability of stem cell subsets in mice

1997

View full text Add to dashboard Cite

Hematopoietic reconsitution after transplantation requires the presence of cells with early and late repopulating ability. To determine the contribution of relatively immature and mature hematopoietic progenitor cells in engraftment, we used a modified rhodamine staining procedure to purify populations of rhodamine 123 (Rho)"ght, Rhod"' and Rho-cells from mouse steady-state bone marrow. Following transplantation of these subsets of stem cells in lethally irradiated mice, marrow repopulating cells were mainly present in the small fraction of Rho-cells, indicating that hematopoietic stem cells have a relatively high expression of P-glycoprotein.Similar cell populations could be purified from cytokine-mobilized blood following treatment with cyclophosphamide and G-CSF. The Rhocell population contained the majority of hematopoietic stem cells with in vivo marrow repopulating ability. In methylcellulose colony assays, the majority of the committed progenitor cells were present in Rhod"" and Rhohrigh' fractions. These results indicate that relatively primitive populations of hematopoietic stem cells that lack colony-forming capacity in vitro mediate the early phase of engraftment following syngeneic stem cell transplantation. Stem Cells 1997;15(suppl 1):47-53

show abstract

Section: Bone Marrow Repopulation Following Syngeneic Transplantationcontrasting

confidence: 85%

Differential short&hyphen;term and long&hyphen;term repopulating ability of stem cell subsets in mice

1997

View full text Add to dashboard Cite

show abstract

“…However, Rho high cells perform poorly in transplantation experiments, producing mainly erythrocytes and contributing little to platelet or granulocyte recovery. 5,6 All previous studies of the Rho low and Rho high subsets have tracked leukocyte engraftment, using mice allelic at the CD45 locus to follow donor-and host-derived hematopoiesis. [5][6][7][8] Although extremely useful, this model does not allow for tracking of erythroid or platelet engraftment, because these lineages lack CD45.…”

Section: Introductionmentioning

confidence: 99%

The spleen is a major site of megakaryopoiesis following transplantation of murine hematopoietic stem cells

Slayton

Georgelas

Pierce

et al. 2002

Blood

View full text Add to dashboard Cite

The stem cell pool can be fractionated by using the mitochondrial dye, rhodamine-123, into Rho low hematopoietic stem cells and Rho high progenitors. Rho low stem cells permanently engraft all lineages, whereas Rho high progenitors transiently produce erythrocytes, without substantial platelet or granulocyte production. We hypothesized that the inability of the Rho high cells to produce platelets in vivo was due to the fact that these cells preferentially engraft in the spleen and lack marrow engraftment. Initially, we demonstrated that Rho high progenitors produced more megakaryocytes in vitro than Rho low stem cells did. To study the activity of the Rho low and Rho high subsets in vivo, we used mice allelic at the hemoglobin and glucose phosphate isomerase loci to track donor-derived erythropoiesis and thrombopoiesis. Rho low stem cells contributed to robust and long-term erythroid and platelet engraftment, whereas Rho high progenitors contributed only to transient erythroid engraftment and produced very low numbers of platelets in vivo. Donorderived megakaryopoiesis occurred at higher densities in the spleen than in the bone marrow in animals receiving Rho low stem cells and peaked around day 28. Blockade of splenic engraftment using pertussis toxin did not affect the peak of splenic megakaryopoiesis, supporting the hypothesis that these megakaryocytes were derived from progenitors that originated in the bone marrow. These data emphasize that in vitro behavior of hematopoietic progenitor cell subsets does not always predict their behavior following transplantation. This study supports a major role for the spleen in thrombopoi IntroductionMurine hematopoietic stem cells have been shown to reside within a population of cells that do not express lineage-specific markers (Lin neg ) and express the antigens Sca-1, c-kit, and Thy-1.1. Cells of this phenotype comprise the stem cell pool in certain strains of mice. 1,2 Rhodamine-123 (Rho) is a mitochondrial dye that stains cells based on their state of activation in which the more metabolically active cells tend to fluoresce brightly and quiescent cells dimly. 3 Rho has been used to subfractionate the stem cell pool into quiescent and metabolically active subsets. The Rho low subset is highly enriched for hematopoietic stem cells, to the degree that as few as 4 of these cells can reconstitute a mouse for the lifetime of the animal. Observations of the behavior of these stem cells have led to the model of the 2-tiered stem-cell pool. 4 According to this model, the Rho low cells make rare contributions to the peripheral compartment, but are mobilized in response to physiologic needs, and represent most of the transplantable stem cells. In contrast, Rho high cells have a high rate of cell division and, thus, make frequent contributions to the peripheral compartment under normal conditions. However, Rho high cells perform poorly in transplantation experiments, producing mainly erythrocytes and contributing little to platelet or granulocyte recovery. 5,6 All previous stu...

show abstract

“…However, H-2 antibody typing of spleen cells is restricted to MHC-mismatched allogeneic BMT where host-versus-graft and graft-versus-host reactions influence BM engraftment. Flow cytometric analysis of other surface markers (Ly-1, Ly-5, and Thy-1) was developed as a method for determination of long-term chimerism in congenic recipients [14,15]. Ly-1 and Thy-1 markers can only be used for measurement of lymphoid chimerism and the Ly-5 antigen (CD45) is expressed on all BM-derived cells except erythrocytes and erythroblasts [16].…”

Section: Introductionmentioning

confidence: 99%

Immunogenicity of Ly5 (CD45)‐Antigens Hampers Long‐Term Engraftment Following Minimal Conditioning in a Murine Bone Marrow Transplantation Model

Sheridan

Robinson

et al. 2001

STEM CELLS

View full text Add to dashboard Cite

Various techniques are available for distinguishing donor from host cells evaluating the efficacy of conditioning regimen for experimental bone marrow transplantation (BMT). Techniques include the use of extracellular immunological markers, such as Ly5 (CD45), and intracellular biochemical markers, such as glucose-phosphate-isomerase (Gpi). Because Ly5 is an extracellular protein, the disparity between donor (Ly5.1) and host (Ly5.2) antigens may induce a weak immune response whereas with Gpi, no immune response is expected. This difference may be of particular concern in experimental transplantation approaches that use minimal conditioning such as low-dose total body irradiation (TBI). Such mild conditioning may not induce the immunosuppression required to overcome host rejection of Ly5 disparate cells.To ). However, when higher TBI or BMC doses were used, similar engraftment levels were found, suggesting sufficient immune suppression to allow equal engraftment of both sources of BM.These data suggest that even a minor phenotypic disparity between donor and host, such as Ly5, may necessitate high-dose TBI to prevent rejection. The combination of low-dose TBI or other nonmyeloablative conditioning strategies with small numbers of BMC may lead to reduced engraftment when extracellular immunological markers such as Ly5 are used for transplantation studies. Therefore, small immunological differences must be considered when using the Ly5 marker for engraftment.

show abstract

Rhodamine123 reveals heterogeneity within murine Lin-, Sca-1+ hemopoietic stem cells.

Cited by 149 publications

References 10 publications

Differential short&hyphen;term and long&hyphen;term repopulating ability of stem cell subsets in mice

Differential short&hyphen;term and long&hyphen;term repopulating ability of stem cell subsets in mice

The spleen is a major site of megakaryopoiesis following transplantation of murine hematopoietic stem cells

Immunogenicity of Ly5 (CD45)‐Antigens Hampers Long‐Term Engraftment Following Minimal Conditioning in a Murine Bone Marrow Transplantation Model

Contact Info

Product

Resources

About