Long-lived intragraft donor leukocytes or relocated donor hematopoietic stem/progenitor cells can cause long-term hematopoietic chimerism after liver transplantation

Abstract Cryopreservation of hematopoietic stem cells (HSCs) and hematopoietic progenitor cells (HPCs) is crucial for cord blood (CB) banking and transplantation. We evaluated recovery of functional HPC cryopreserved as mononuclear or unseparated cells for up to 23.5 years compared with prefreeze values of the same CB units. Highly efficient recovery (80%-100%) was apparent for granulocyte-macrophage and multipotential hematopoietic progenitors, although some collections had reproducible low recovery. Proliferative potential, response to multiple cytokines, and replating of HPC colonies was extensive. CD34+ cells isolated from CB cryopreserved for up to 21 years had long-term (≥ 6 month) engrafting capability in primary and secondary immunodeficient mice reflecting recovery of long-term repopulating, self-renewing HSCs. We recovered functionally responsive CD4+ and CD8+ T lymphocytes, generated induced pluripotent stem (iPS) cells with differentiation representing all 3 germ cell lineages in vitro and in vivo, and detected high proliferative endothelial colony forming cells, results of relevance to CB biology and banking.

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Hematopoietic stem cells in the blood after stem cell factor and interleukin-11 administration: evidence for different mechanisms of mobilization

Blood ◽

10.1182/blood.v86.12.4674.bloodjournal86124674 ◽

1995 ◽

Vol 86 (12) ◽

pp. 4674-4680 ◽

Cited By ~ 35

Author(s):

P Mauch ◽

C Lamont ◽

TY Neben ◽

C Quinto ◽

SJ Goldman ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Progenitor Cells ◽

Stem Cell Factor ◽

Cytotoxic Agents ◽

Peripheral Blood Stem Cells ◽

Hematopoietic Stem ◽

Blood Stem Cells ◽

Interleukin 11

Peripheral blood stem cells and progenitor cells, collected during recovery from exposure to cytotoxic agents or after cytokine administration, are being increasingly used in clinical bone marrow transplantation. To determine factors important for mobilization of both primitive stem cells and progenitor cells to the blood, we studied the blood and splenic and marrow compartments of intact and splenectomized mice after administration of recombinant human interleukin-11 (rhlL-11), recombinant rat stem cell factor (rrSCF), and IL-11 + SCF. IL-11 administration increased the number of spleen colony- forming units (CFU-S) in both the spleen and blood, but did not increase blood long-term marrow-repopulating ability (LTRA) in intact or splenectomized mice. SCF administration increased the number of CFU- S in both the spleen and blood and did not increase the blood or splenic LTRA of intact mice, but did increase blood LTRA to normal marrow levels in splenectomized mice. The combination of lL-11 + SCF syngeristically enhanced mobilization of long-term marrow-repopulating cells from the marrow to the spleen of intact mice and from the marrow to the blood of splenectomized mice. These data, combined with those of prior studies showing granulocyte colony-stimulating factor mobilization of long-term marrow repopulating cells from the marrow to the blood of mice with intact spleens, suggest different cytokine- induced pathways for mobilization of primitive stem cells.

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Adhesion to Fibronectin Maintains Regenerative Capacity During Ex Vivo Culture and Transduction of Human Hematopoietic Stem and Progenitor Cells

Blood ◽

10.1182/blood.v92.12.4612 ◽

1998 ◽

Vol 92 (12) ◽

pp. 4612-4621 ◽

Cited By ~ 128

Author(s):

M.A. Dao ◽

K. Hashino ◽

I. Kato ◽

J.A. Nolta

Keyword(s):

Progenitor Cells ◽

Ex Vivo ◽

Xenograft Model ◽

Retroviral Vectors ◽

Current Data ◽

Regenerative Capacity ◽

Hematopoietic Stem ◽

Stem And Progenitor Cells ◽

Ex Vivo Culture

Abstract Recent reports have indicated that there is poor engraftment from hematopoietic stem cells (HSC) that have traversed cell cycle ex vivo. However, inducing cells to cycle in culture is critical to the fields of ex vivo stem cell expansion and retroviral-mediated gene therapy. Through the use of a xenograft model, the current data shows that human hematopoietic stem and progenitor cells can traverse M phase ex vivo, integrate retroviral vectors, engraft, and sustain long-term hematopoiesis only if they have had the opportunity to engage their integrin receptors to fibronectin during the culture period. If cultured in suspension under the same conditions, transduction is undetectable and the long-term multilineage regenerative capacity of the primitive cells is severely diminished.

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Down-regulation of GATA1 uncouples STAT5-induced erythroid differentiation from stem/progenitor cell proliferation

Blood ◽

10.1182/blood-2009-10-250894 ◽

2010 ◽

Vol 115 (22) ◽

pp. 4367-4376 ◽

Cited By ~ 16

Author(s):

Albertus T. J. Wierenga ◽

Edo Vellenga ◽

Jan Jacob Schuringa

Keyword(s):

Progenitor Cells ◽

Progenitor Cell ◽

Target Genes ◽

Lentiviral Vector ◽

Erythroid Differentiation ◽

Down Regulation ◽

Hematopoietic Stem ◽

Progenitor Cell Proliferation ◽

Cord Blood Cells

Abstract Previously, we have shown that overexpression of an activated mutant of signal transducer and activator of transcription-5 (STAT5) induces erythropoiesis, impaired myelopoiesis, and an increase in long-term proliferation of human hematopoietic stem/progenitor cells. Because GATA1 is a key transcription factor involved in erythropoiesis, the involvement of GATA1 in STAT5-induced phenotypes was studied by shRNA-mediated knockdown of GATA1. CD34+ cord blood cells were double transduced with a conditionally active STAT5 mutant and a lentiviral vector expressing a short hairpin against GATA1. Erythropoiesis was completely abolished in the absence of GATA1, indicating that STAT5-induced erythropoiesis is GATA1-dependent. Furthermore, the impaired myelopoiesis in STAT5-transduced cells was restored by GATA1 knockdown. Interestingly, early cobblestone formation was only modestly affected, and long-term growth of STAT5-positive cells was increased in the absence of GATA1, whereby high progenitor numbers were maintained. Thus, GATA1 down-regulation allowed the dissection of STAT5-induced differentiation phenotypes from the effects on long-term expansion of stem/progenitor cells. Gene expression profiling allowed the identification of GATA1-dependent and GATA1-independent STAT5 target genes, and these studies revealed that several proliferation-related genes were up-regulated by STAT5 independent of GATA1, whereas several erythroid differentiation-related genes were found to be GATA1 as well as STAT5 dependent.

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35. Long-Term Engraftment and In Vivo Selection of Hematopoietic Stem/Progenitor Cells Generated By Vascular Niche Induction of Induced Pluripotent Stem Cells

Molecular Therapy ◽

10.1016/s1525-0016(16)35048-1 ◽

2014 ◽

Vol 22 ◽

pp. S14

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Progenitor Cells ◽

Pluripotent Stem Cells ◽

Hematopoietic Stem ◽

In Vivo Selection ◽

Induced Pluripotent ◽

Selection Of

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The AML1-ETO fusion protein promotes the expansion of human hematopoietic stem cells

Blood ◽

10.1182/blood.v99.1.15 ◽

2002 ◽

Vol 99 (1) ◽

pp. 15-23 ◽

Cited By ~ 141

Author(s):

James C. Mulloy ◽

Jörg Cammenga ◽

Karen L. MacKenzie ◽

Francisco J. Berguido ◽

Malcolm A. S. Moore ◽

...

Keyword(s):

Stem Cells ◽

Fusion Protein ◽

Progenitor Cells ◽

Protein Interactions ◽

Dominant Negative ◽

Myelogenous Leukemia ◽

Hematopoietic Stem ◽

Stem And Progenitor Cells

The acute myelogenous leukemia–1 (AML1)–ETO fusion protein is generated by the t(8;21), which is found in 40% of AMLs of the French-American-British M2 subtype. AML1-ETO interferes with the function of the AML1 (RUNX1, CBFA2) transcription factor in a dominant-negative fashion and represses transcription by binding its consensus DNA–binding site and via protein-protein interactions with other transcription factors. AML1 activity is critical for the development of definitive hematopoiesis, and haploinsufficiency of AML1 has been linked to a propensity to develop AML. Murine experiments suggest that AML1-ETO expression may not be sufficient for leukemogenesis; however, like the BCR-ABL isoforms, the cellular background in which these fusion proteins are expressed may be critical to the phenotype observed. Retroviral gene transfer was used to examine the effect of AML1-ETO on the in vitro behavior of human hematopoietic stem and progenitor cells. Following transduction of CD34+ cells, stem and progenitor cells were quantified in clonogenic assays, cytokine-driven expansion cultures, and long-term stromal cocultures. Expression of AML1-ETO inhibited colony formation by committed progenitors, but enhanced the growth of stem cells (cobblestone area-forming cells), resulting in a profound survival advantage of transduced over nontransduced cells. AML1-ETO–expressing cells retained progenitor activity and continued to express CD34 throughout the 5-week long-term culture. Thus, AML1-ETO enhances the self-renewal of pluripotent stem cells, the physiological target of many acute myeloid leukemias.

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