Clinically Relevant Expansion of Hematopoietic Stem Cells with Conserved Function in a Single-Use, Closed-System Bioprocess

The clinical potential of umbilical cord blood-derived stem and progenitor cells has been demonstrated in various animal and human transplantation studies. However, the need for increased numbers of appropriate umbilical cord blood-derived cells continues to limit the development and success of thes...

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Published inBiology of blood and marrow transplantation Vol. 12; no. 10; pp. 1020 - 1030
Main Authors Madlambayan, Gerard J., Rogers, Ian, Purpura, Kelly A., Ito, Caryn, Yu, Mei, Kirouac, Daniel, Casper, Robert F., Zandstra, Peter W.
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.10.2006
Subjects
Animals
B-Lymphocyte Subsets - cytology
B-Lymphocyte Subsets - immunology
Bioprocess development
Cell Culture Techniques - instrumentation
Cell Culture Techniques - methods
Cell Differentiation
Cell Lineage
Cell Separation - methods
Cells, Cultured - cytology
Colony-Forming Units Assay
Cord blood
Cord Blood Stem Cell Transplantation - methods
Fetal Blood - cytology
Graft Survival
Hematopoiesis
Hematopoietic Stem Cells - cytology
Humans
Immunophenotyping
Infant, Newborn
Killer Cells, Natural - cytology
Killer Cells, Natural - immunology
Lymphocytes - cytology
Lymphocytes - immunology
Mice
Mice, Inbred NOD
Mice, SCID
Radiation Chimera
Stem cell expansion
T-Lymphocyte Subsets - cytology
T-Lymphocyte Subsets - immunology
Transplantation, Heterologous
Stem cell expansion
Bioprocess development
Cord blood
Hematopoiesis
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Abstract The clinical potential of umbilical cord blood-derived stem and progenitor cells has been demonstrated in various animal and human transplantation studies. However, the need for increased numbers of appropriate umbilical cord blood-derived cells continues to limit the development and success of these therapies. Ex vivo expansion has been widely studied as a method to overcome this limitation. We describe the use of a clinically relevant single-use, closed-system bioprocess capable of generating greater numbers of hematopoietic stem and progenitor cells that maintain in vivo and in vitro developmental potential. In addition to expanded numbers of CD34 + cells, CD34 +CD38 − cells, colony-forming cells, and long-term culture-initiating cells, the bioprocess generated ≥3.3-fold more long-term nonobese diabetic/severe combined immunodeficient repopulating cells (quantitatively determined using limiting dilution analysis) than present at input. Interestingly, these cells were also capable of multilineage engraftment and were shown to maintain their engraftment potency on a per long-term nonobese diabetic/severe combined immunodeficient repopulating cell basis compared with input noncultured cells. The developmental capacity of bioprocess-generated cells was further demonstrated by their ability to repopulate secondary nonobese diabetic/severe combined immunodeficient recipients. In vitro lineage analysis confirmed that bioprocess-generated cells could differentiate into myeloid and natural killer, B, and T cell lymphoid lineages. This in-depth analysis describes a bioprocess that generates human hematopoietic stem and progenitor cells with conserved hematopoietic activity, establishes analysis criteria for in vitro hematopoietic stem cell expansion studies, and serves as a foundation to test the therapeutic utility of cultured hematopoietic stem cells in large animals and humans.
AbstractList The clinical potential of umbilical cord blood-derived stem and progenitor cells has been demonstrated in various animal and human transplantation studies. However, the need for increased numbers of appropriate umbilical cord blood-derived cells continues to limit the development and success of these therapies. Ex vivo expansion has been widely studied as a method to overcome this limitation. We describe the use of a clinically relevant single-use, closed-system bioprocess capable of generating greater numbers of hematopoietic stem and progenitor cells that maintain in vivo and in vitro developmental potential. In addition to expanded numbers of CD34+ cells, CD34(+)CD38(-) cells, colony-forming cells, and long-term culture-initiating cells, the bioprocess generated > or =3.3-fold more long-term nonobese diabetic/severe combined immunodeficient repopulating cells (quantitatively determined using limiting dilution analysis) than present at input. Interestingly, these cells were also capable of multilineage engraftment and were shown to maintain their engraftment potency on a per long-term nonobese diabetic/severe combined immunodeficient repopulating cell basis compared with input noncultured cells. The developmental capacity of bioprocess-generated cells was further demonstrated by their ability to repopulate secondary nonobese diabetic/severe combined immunodeficient recipients. In vitro lineage analysis confirmed that bioprocess-generated cells could differentiate into myeloid and natural killer, B, and T cell lymphoid lineages. This in-depth analysis describes a bioprocess that generates human hematopoietic stem and progenitor cells with conserved hematopoietic activity, establishes analysis criteria for in vitro hematopoietic stem cell expansion studies, and serves as a foundation to test the therapeutic utility of cultured hematopoietic stem cells in large animals and humans.
The clinical potential of umbilical cord blood-derived stem and progenitor cells has been demonstrated in various animal and human transplantation studies. However, the need for increased numbers of appropriate umbilical cord blood-derived cells continues to limit the development and success of these therapies. Ex vivo expansion has been widely studied as a method to overcome this limitation. We describe the use of a clinically relevant single-use, closed-system bioprocess capable of generating greater numbers of hematopoietic stem and progenitor cells that maintain in vivo and in vitro developmental potential. In addition to expanded numbers of CD34 + cells, CD34 +CD38 − cells, colony-forming cells, and long-term culture-initiating cells, the bioprocess generated ≥3.3-fold more long-term nonobese diabetic/severe combined immunodeficient repopulating cells (quantitatively determined using limiting dilution analysis) than present at input. Interestingly, these cells were also capable of multilineage engraftment and were shown to maintain their engraftment potency on a per long-term nonobese diabetic/severe combined immunodeficient repopulating cell basis compared with input noncultured cells. The developmental capacity of bioprocess-generated cells was further demonstrated by their ability to repopulate secondary nonobese diabetic/severe combined immunodeficient recipients. In vitro lineage analysis confirmed that bioprocess-generated cells could differentiate into myeloid and natural killer, B, and T cell lymphoid lineages. This in-depth analysis describes a bioprocess that generates human hematopoietic stem and progenitor cells with conserved hematopoietic activity, establishes analysis criteria for in vitro hematopoietic stem cell expansion studies, and serves as a foundation to test the therapeutic utility of cultured hematopoietic stem cells in large animals and humans.
The clinical potential of umbilical cord blood-derived stem and progenitor cells has been demonstrated in various animal and human transplantation studies. However, the need for increased numbers of appropriate umbilical cord blood-derived cells continues to limit the development and success of these therapies. Ex vivo expansion has been widely studied as a method to overcome this limitation. We describe the use of a clinically relevant single-use, closed-system bioprocess capable of generating greater numbers of hematopoietic stem and progenitor cells that maintain in vivo and in vitro developmental potential. In addition to expanded numbers of CD34+ cells, CD34(+)CD38(-) cells, colony-forming cells, and long-term culture-initiating cells, the bioprocess generated > or =3.3-fold more long-term nonobese diabetic/severe combined immunodeficient repopulating cells (quantitatively determined using limiting dilution analysis) than present at input. Interestingly, these cells were also capable of multilineage engraftment and were shown to maintain their engraftment potency on a per long-term nonobese diabetic/severe combined immunodeficient repopulating cell basis compared with input noncultured cells. The developmental capacity of bioprocess-generated cells was further demonstrated by their ability to repopulate secondary nonobese diabetic/severe combined immunodeficient recipients. In vitro lineage analysis confirmed that bioprocess-generated cells could differentiate into myeloid and natural killer, B, and T cell lymphoid lineages. This in-depth analysis describes a bioprocess that generates human hematopoietic stem and progenitor cells with conserved hematopoietic activity, establishes analysis criteria for in vitro hematopoietic stem cell expansion studies, and serves as a foundation to test the therapeutic utility of cultured hematopoietic stem cells in large animals and humans.
Author Ito, Caryn
Kirouac, Daniel
Purpura, Kelly A.
Casper, Robert F.
Madlambayan, Gerard J.
Yu, Mei
Rogers, Ian
Zandstra, Peter W.
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  fullname: Zandstra, Peter W.
  email: peter.zandstra@utoronto.ca
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/17084368$$D View this record in MEDLINE/PubMed
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Issue 10
Keywords Stem cell expansion
Bioprocess development
Cord blood
Hematopoiesis
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Snippet The clinical potential of umbilical cord blood-derived stem and progenitor cells has been demonstrated in various animal and human transplantation studies....
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SubjectTerms Animals
B-Lymphocyte Subsets - cytology
B-Lymphocyte Subsets - immunology
Bioprocess development
Cell Culture Techniques - instrumentation
Cell Culture Techniques - methods
Cell Differentiation
Cell Lineage
Cell Separation - methods
Cells, Cultured - cytology
Colony-Forming Units Assay
Cord blood
Cord Blood Stem Cell Transplantation - methods
Fetal Blood - cytology
Graft Survival
Hematopoiesis
Hematopoietic Stem Cells - cytology
Humans
Immunophenotyping
Infant, Newborn
Killer Cells, Natural - cytology
Killer Cells, Natural - immunology
Lymphocytes - cytology
Lymphocytes - immunology
Mice
Mice, Inbred NOD
Mice, SCID
Radiation Chimera
Stem cell expansion
T-Lymphocyte Subsets - cytology
T-Lymphocyte Subsets - immunology
Transplantation, Heterologous
Title Clinically Relevant Expansion of Hematopoietic Stem Cells with Conserved Function in a Single-Use, Closed-System Bioprocess
URI https://dx.doi.org/10.1016/j.bbmt.2006.07.005
https://www.ncbi.nlm.nih.gov/pubmed/17084368
https://search.proquest.com/docview/68114796
Volume 12
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