Expansion and preservation of the functional activity of adult hematopoietic stem cells cultured ex vivo with a histone deacetylase inhibitor
Attempts to expand ex vivo the numbers of human hematopoietic stem cells (HSCs) without compromising their marrow repopulating capacity and their ability to establish multilineage hematopoiesis has been the subject of intense investigation. Although most such efforts have focused on cord blood HSCs,...
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| Published in | Stem cells translational medicine Vol. 9; no. 4; pp. 531 - 542 |
|---|---|
| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Hoboken, USA
John Wiley & Sons, Inc
01.04.2020
Oxford University Press |
| Subjects | |
| Online Access | Get full text |
| ISSN | 2157-6564 2157-6580 2157-6580 |
| DOI | 10.1002/sctm.19-0199 |
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| Abstract | Attempts to expand ex vivo the numbers of human hematopoietic stem cells (HSCs) without compromising their marrow repopulating capacity and their ability to establish multilineage hematopoiesis has been the subject of intense investigation. Although most such efforts have focused on cord blood HSCs, few have been applied to adult HSCs, a more clinically relevant HSC source for gene modification. To date, the strategies that have been used to expand adult HSCs have resulted in modest effects or HSCs with lineage bias and a limited ability to generate T cells in vivo. We previously reported that culturing umbilical cord blood CD34+ cells in serum‐free media supplemented with valproic acid (VPA), a histone deacetylase inhibitor, and a combination of cytokines led to the expansion of the numbers of fully functional HSCs. In the present study, we used this same approach to expand the numbers of adult human CD34+ cells isolated from mobilized peripheral blood and bone marrow. This approach resulted in a significant increase in the numbers of phenotypically defined HSCs (CD34+CD45RA‐CD90+D49f+). Cells incubated with VPA also exhibited increased aldehyde dehydrogenase activity and decreased mitochondrial membrane potential, each functional markers of HSCs. Grafts harvested from VPA‐treated cultures were able to engraft in immune‐deficient mice and, importantly, to generate cellular progeny belonging to each hematopoietic lineage in similar proportion to that observed with unmanipulated CD34+ cells. These data support the utility of VPA‐mediated ex vivo HSC expansion for gene modification of adult HSCs.
Valproic acid (VPA)‐mediated ex vivo expansion of adult bone marrow and mobilized peripheral blood CD34+ cells resulted in a cellular product characterized by high viability, enrichment with CD34+CD45RA‐CD90+ cells, increased aldehyde dehydrogenase (ALDH) activity, robust multipotent clonogenic potential, and decreased mitochondrial potential. VPA‐expanded grafts were able to establish unbiased multilineage human hematopoietic‐cell chimerism in NSG mice at 16 weeks post‐transplantation. |
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| AbstractList | Attempts to expand ex vivo the numbers of human hematopoietic stem cells (HSCs) without compromising their marrow repopulating capacity and their ability to establish multilineage hematopoiesis has been the subject of intense investigation. Although most such efforts have focused on cord blood HSCs, few have been applied to adult HSCs, a more clinically relevant HSC source for gene modification. To date, the strategies that have been used to expand adult HSCs have resulted in modest effects or HSCs with lineage bias and a limited ability to generate T cells in vivo. We previously reported that culturing umbilical cord blood CD34+ cells in serum‐free media supplemented with valproic acid (VPA), a histone deacetylase inhibitor, and a combination of cytokines led to the expansion of the numbers of fully functional HSCs. In the present study, we used this same approach to expand the numbers of adult human CD34+ cells isolated from mobilized peripheral blood and bone marrow. This approach resulted in a significant increase in the numbers of phenotypically defined HSCs (CD34+CD45RA‐CD90+D49f+). Cells incubated with VPA also exhibited increased aldehyde dehydrogenase activity and decreased mitochondrial membrane potential, each functional markers of HSCs. Grafts harvested from VPA‐treated cultures were able to engraft in immune‐deficient mice and, importantly, to generate cellular progeny belonging to each hematopoietic lineage in similar proportion to that observed with unmanipulated CD34+ cells. These data support the utility of VPA‐mediated ex vivo HSC expansion for gene modification of adult HSCs. Valproic acid (VPA)‐mediated ex vivo expansion of adult bone marrow and mobilized peripheral blood CD34+ cells resulted in a cellular product characterized by high viability, enrichment with CD34+CD45RA‐CD90+ cells, increased aldehyde dehydrogenase (ALDH) activity, robust multipotent clonogenic potential, and decreased mitochondrial potential. VPA‐expanded grafts were able to establish unbiased multilineage human hematopoietic‐cell chimerism in NSG mice at 16 weeks post‐transplantation. Attempts to expand ex vivo the numbers of human hematopoietic stem cells (HSCs) without compromising their marrow repopulating capacity and their ability to establish multilineage hematopoiesis has been the subject of intense investigation. Although most such efforts have focused on cord blood HSCs, few have been applied to adult HSCs, a more clinically relevant HSC source for gene modification. To date, the strategies that have been used to expand adult HSCs have resulted in modest effects or HSCs with lineage bias and a limited ability to generate T cells in vivo. We previously reported that culturing umbilical cord blood CD34+ cells in serum-free media supplemented with valproic acid (VPA), a histone deacetylase inhibitor, and a combination of cytokines led to the expansion of the numbers of fully functional HSCs. In the present study, we used this same approach to expand the numbers of adult human CD34+ cells isolated from mobilized peripheral blood and bone marrow. This approach resulted in a significant increase in the numbers of phenotypically defined HSCs (CD34+CD45RA-CD90+D49f+). Cells incubated with VPA also exhibited increased aldehyde dehydrogenase activity and decreased mitochondrial membrane potential, each functional markers of HSCs. Grafts harvested from VPA-treated cultures were able to engraft in immune-deficient mice and, importantly, to generate cellular progeny belonging to each hematopoietic lineage in similar proportion to that observed with unmanipulated CD34+ cells. These data support the utility of VPA-mediated ex vivo HSC expansion for gene modification of adult HSCs. Attempts to expand ex vivo the numbers of human hematopoietic stem cells (HSCs) without compromising their marrow repopulating capacity and their ability to establish multilineage hematopoiesis has been the subject of intense investigation. Although most such efforts have focused on cord blood HSCs, few have been applied to adult HSCs, a more clinically relevant HSC source for gene modification. To date, the strategies that have been used to expand adult HSCs have resulted in modest effects or HSCs with lineage bias and a limited ability to generate T cells in vivo. We previously reported that culturing umbilical cord blood CD34+ cells in serum-free media supplemented with valproic acid (VPA), a histone deacetylase inhibitor, and a combination of cytokines led to the expansion of the numbers of fully functional HSCs. In the present study, we used this same approach to expand the numbers of adult human CD34+ cells isolated from mobilized peripheral blood and bone marrow. This approach resulted in a significant increase in the numbers of phenotypically defined HSCs (CD34+CD45RA-CD90+D49f+). Cells incubated with VPA also exhibited increased aldehyde dehydrogenase activity and decreased mitochondrial membrane potential, each functional markers of HSCs. Grafts harvested from VPA-treated cultures were able to engraft in immune-deficient mice and, importantly, to generate cellular progeny belonging to each hematopoietic lineage in similar proportion to that observed with unmanipulated CD34+ cells. These data support the utility of VPA-mediated ex vivo HSC expansion for gene modification of adult HSCs.Attempts to expand ex vivo the numbers of human hematopoietic stem cells (HSCs) without compromising their marrow repopulating capacity and their ability to establish multilineage hematopoiesis has been the subject of intense investigation. Although most such efforts have focused on cord blood HSCs, few have been applied to adult HSCs, a more clinically relevant HSC source for gene modification. To date, the strategies that have been used to expand adult HSCs have resulted in modest effects or HSCs with lineage bias and a limited ability to generate T cells in vivo. We previously reported that culturing umbilical cord blood CD34+ cells in serum-free media supplemented with valproic acid (VPA), a histone deacetylase inhibitor, and a combination of cytokines led to the expansion of the numbers of fully functional HSCs. In the present study, we used this same approach to expand the numbers of adult human CD34+ cells isolated from mobilized peripheral blood and bone marrow. This approach resulted in a significant increase in the numbers of phenotypically defined HSCs (CD34+CD45RA-CD90+D49f+). Cells incubated with VPA also exhibited increased aldehyde dehydrogenase activity and decreased mitochondrial membrane potential, each functional markers of HSCs. Grafts harvested from VPA-treated cultures were able to engraft in immune-deficient mice and, importantly, to generate cellular progeny belonging to each hematopoietic lineage in similar proportion to that observed with unmanipulated CD34+ cells. These data support the utility of VPA-mediated ex vivo HSC expansion for gene modification of adult HSCs. Attempts to expand ex vivo the numbers of human hematopoietic stem cells (HSCs) without compromising their marrow repopulating capacity and their ability to establish multilineage hematopoiesis has been the subject of intense investigation. Although most such efforts have focused on cord blood HSCs, few have been applied to adult HSCs, a more clinically relevant HSC source for gene modification. To date, the strategies that have been used to expand adult HSCs have resulted in modest effects or HSCs with lineage bias and a limited ability to generate T cells in vivo. We previously reported that culturing umbilical cord blood CD34+ cells in serum‐free media supplemented with valproic acid (VPA), a histone deacetylase inhibitor, and a combination of cytokines led to the expansion of the numbers of fully functional HSCs. In the present study, we used this same approach to expand the numbers of adult human CD34+ cells isolated from mobilized peripheral blood and bone marrow. This approach resulted in a significant increase in the numbers of phenotypically defined HSCs (CD34+CD45RA‐CD90+D49f+). Cells incubated with VPA also exhibited increased aldehyde dehydrogenase activity and decreased mitochondrial membrane potential, each functional markers of HSCs. Grafts harvested from VPA‐treated cultures were able to engraft in immune‐deficient mice and, importantly, to generate cellular progeny belonging to each hematopoietic lineage in similar proportion to that observed with unmanipulated CD34+ cells. These data support the utility of VPA‐mediated ex vivo HSC expansion for gene modification of adult HSCs. Valproic acid (VPA)‐mediated ex vivo expansion of adult bone marrow and mobilized peripheral blood CD34+ cells resulted in a cellular product characterized by high viability, enrichment with CD34+CD45RA‐CD90+ cells, increased aldehyde dehydrogenase (ALDH) activity, robust multipotent clonogenic potential, and decreased mitochondrial potential. VPA‐expanded grafts were able to establish unbiased multilineage human hematopoietic‐cell chimerism in NSG mice at 16 weeks post‐transplantation. Abstract Attempts to expand ex vivo the numbers of human hematopoietic stem cells (HSCs) without compromising their marrow repopulating capacity and their ability to establish multilineage hematopoiesis has been the subject of intense investigation. Although most such efforts have focused on cord blood HSCs, few have been applied to adult HSCs, a more clinically relevant HSC source for gene modification. To date, the strategies that have been used to expand adult HSCs have resulted in modest effects or HSCs with lineage bias and a limited ability to generate T cells in vivo. We previously reported that culturing umbilical cord blood CD34+ cells in serum‐free media supplemented with valproic acid (VPA), a histone deacetylase inhibitor, and a combination of cytokines led to the expansion of the numbers of fully functional HSCs. In the present study, we used this same approach to expand the numbers of adult human CD34+ cells isolated from mobilized peripheral blood and bone marrow. This approach resulted in a significant increase in the numbers of phenotypically defined HSCs (CD34+CD45RA‐CD90+D49f+). Cells incubated with VPA also exhibited increased aldehyde dehydrogenase activity and decreased mitochondrial membrane potential, each functional markers of HSCs. Grafts harvested from VPA‐treated cultures were able to engraft in immune‐deficient mice and, importantly, to generate cellular progeny belonging to each hematopoietic lineage in similar proportion to that observed with unmanipulated CD34+ cells. These data support the utility of VPA‐mediated ex vivo HSC expansion for gene modification of adult HSCs. |
| Audience | Academic |
| Author | Papa, Luena Patel, Ami Iancu‐Rubin, Camelia Djedaini, Mansour Zimran, Eran Hoffman, Ronald |
| AuthorAffiliation | 1 Division of Hematology and Oncology Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai New York NY 2 Hematology Department Hadassah University Center Jerusalem Israel |
| AuthorAffiliation_xml | – name: 2 Hematology Department Hadassah University Center Jerusalem Israel – name: 1 Division of Hematology and Oncology Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai New York NY |
| Author_xml | – sequence: 1 givenname: Eran orcidid: 0000-0002-5483-5180 surname: Zimran fullname: Zimran, Eran email: eran.zimran@gmail.com organization: Hadassah University Center – sequence: 2 givenname: Luena surname: Papa fullname: Papa, Luena organization: Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai – sequence: 3 givenname: Mansour surname: Djedaini fullname: Djedaini, Mansour organization: Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai – sequence: 4 givenname: Ami surname: Patel fullname: Patel, Ami organization: Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai – sequence: 5 givenname: Camelia surname: Iancu‐Rubin fullname: Iancu‐Rubin, Camelia organization: Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai – sequence: 6 givenname: Ronald surname: Hoffman fullname: Hoffman, Ronald organization: Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/31950644$$D View this record in MEDLINE/PubMed |
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| Keywords | adult bone marrow valproic acid hematopoietic stem cells mobilized peripheral blood ex vivo expansion histone deacetylase inhibitor |
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start-page: 864 year: 2015 ident: 2022022809302389200_sct312661-bib-0023 article-title: Ex vivo expansion of human mobilized peripheral blood stem cells using epigenetic modifiers publication-title: Transfusion doi: 10.1111/trf.12904 – volume: 21 start-page: 2581 issue: 4 year: 2012 ident: 2022022809302389200_sct312661-bib-0034 article-title: Histone deacetylase 3 modulates the expansion of human hematopoietic stem cells publication-title: Stem Cells Dev doi: 10.1089/scd.2011.0698 – volume: 130 start-page: 3341 year: 2017 ident: 2022022809302389200_sct312661-bib-0025 article-title: Aryl hydrocarbon receptor antagonists expand adult hematopoietic stem cells from mobilized peripheral blood and bone marrow and increase the dose of CRISPR/Cas9 gene-edited NSG-repopulating cells publication-title: Blood |
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| Snippet | Attempts to expand ex vivo the numbers of human hematopoietic stem cells (HSCs) without compromising their marrow repopulating capacity and their ability to... Abstract Attempts to expand ex vivo the numbers of human hematopoietic stem cells (HSCs) without compromising their marrow repopulating capacity and their... |
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| SubjectTerms | adult bone marrow Aldehyde dehydrogenase Blood Bone marrow CD34 antigen CD45RA antigen CD90 antigen Cord blood Cytokines Divalproex Ethylenediaminetetraacetic acid ex vivo expansion Expansion Experiments Flow cytometry Genetically modified organisms Granulocytes Hematology Hematopoietic stem cells Histone deacetylase histone deacetylase inhibitor Kinases Laboratories Ligands Lymphocytes T Membrane potential Mitochondria mobilized peripheral blood Ontology Peripheral blood Stem cell transplantation Stem cells T cells Tissue‐specific Progenitor and Stem Cells Umbilical cord Valproic acid |
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| Title | Expansion and preservation of the functional activity of adult hematopoietic stem cells cultured ex vivo with a histone deacetylase inhibitor |
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