Co‐culture with mesenchymal stromal cells increases proliferation and maintenance of haematopoietic progenitor cells

Mesenchymal stromal cells (MSC) have been suggested to provide a suitable cellular environment for in vitro expansion of haematopoietic stem and progenitor cells (HPC) from umbilical cord blood. In this study, we have simultaneously analysed the cell division history and immunophenotypic differentia...

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Published inJournal of cellular and molecular medicine Vol. 14; no. 1‐2; pp. 337 - 350
Main Authors Walenda, Thomas, Bork, Simone, Horn, Patrick, Wein, Frederik, Saffrich, Rainer, Diehlmann, Anke, Eckstein, Volker, Ho, Anthony D., Wagner, Wolfgang
Format Journal Article
LanguageEnglish
Published Oxford, UK Blackwell Publishing Ltd 01.01.2010
John Wiley & Sons, Inc
Subjects
adhesion proteins
ADP-ribosyl Cyclase 1 - metabolism
Antibodies
Antigens, CD34 - metabolism
Biotechnology
Carboxyfluorescein diacetate
CD13 antigen
CD34 antigen
CD38 antigen
CD44 antigen
CD45 antigen
CD56 antigen
Cell Adhesion Molecules - genetics
Cell Adhesion Molecules - metabolism
Cell culture
Cell Culture Techniques
Cell differentiation
Cell Differentiation - physiology
Cell division
Cell Proliferation
Cell self-renewal
Cells, Cultured
Cellular Senescence - physiology
Cloning
Coculture Techniques
Cord blood
co‐culture
Fetal Blood - cytology
Flow cytometry
Genetics
Genotype & phenotype
haematopoietic stem cells
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - physiology
Hemopoiesis
Human subjects
Humans
immunophenotype
Immunophenotyping
Kinases
Laboratories
MAP kinase
Mesenchymal stem cells
mesenchymal stromal cells
Mesenchymal Stromal Cells - cytology
Mesenchymal Stromal Cells - physiology
Mitogen-Activated Protein Kinase 1 - antagonists & inhibitors
Mitogen-Activated Protein Kinase 1 - genetics
Mitogen-Activated Protein Kinase 1 - metabolism
N-Cadherin
Progenitor cells
proliferation
Proteins
replicative senescence
RNA, Small Interfering - genetics
RNA, Small Interfering - metabolism
siRNA
stem cell niche
Stem cells
Stromal cells
Stromal Cells - cytology
Stromal Cells - physiology
Studies
Umbilical cord
United States > US
Germany
Online AccessGet full text

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Abstract Mesenchymal stromal cells (MSC) have been suggested to provide a suitable cellular environment for in vitro expansion of haematopoietic stem and progenitor cells (HPC) from umbilical cord blood. In this study, we have simultaneously analysed the cell division history and immunophenotypic differentiation of HPC by using cell division tracking with carboxyfluorescein diacetate N‐succinimidyl ester (CFSE). Co‐culture with MSC greatly enhanced proliferation of human HPC, especially of the more primitive CD34+CD38− fraction. Without co‐culture CD34 and CD133 expressions decreased after several cell divisions, whereas CD38 expression was up‐regulated after some cell divisions and then diminished in fast proliferating cells. Co‐culture with MSC maintained a primitive immunophenotype (CD34+, CD133+ and CD38−) for more population doublings, whereas up‐regulation of differentiation markers (CD13, CD45 and CD56) in HPC was delayed to higher numbers of cell divisions. Especially MSC of early cell passages maintained CD34 expression in HPC over more cell divisions, whereas MSC of higher passages further enhanced their proliferation rate. Inhibition of mitogen‐activated protein kinase 1 (MAPK1) impaired proliferation and differentiation of HPC, but not maintenance of long‐term culture initiating cells. siRNA knockdown of N‐cadherin and VCAM1 in feeder layer cells increased the fraction of slow dividing HPC, whereas knockdown of integrin beta 1 (ITGB1) and CD44 impaired their differentiation. In conclusion, MSC support proliferation as well as self‐renewal of HPC with primitive immunophenotype. The use of early passages of MSC and genetic manipulation of proteins involved in HPC–MSC interaction might further enhance cord blood expansion on MSC.
AbstractList Mesenchymal stromal cells (MSC) have been suggested to provide a suitable cellular environment for in vitro expansion of haematopoietic stem and progenitor cells (HPC) from umbilical cord blood. In this study, we have simultaneously analysed the cell division history and immunophenotypic differentiation of HPC by using cell division tracking with carboxyfluorescein diacetate N-succinimidyl ester (CFSE). Co-culture with MSC greatly enhanced proliferation of human HPC, especially of the more primitive CD34...CD38... fraction. Without co-culture CD34 and CD133 expressions decreased after several cell divisions, whereas CD38 expression was up-regulated after some cell divisions and then diminished in fast proliferating cells. Coculture with MSC maintained a primitive immunophenotype (CD34..., CD133... and CD38-) for more population doublings, whereas up-regulation of differentiation markers (CD13, CD45 and CD56) in HPC was delayed to higher numbers of cell divisions. Especially MSC of early cell passages maintained CD34 expression in HPC over more cell divisions, whereas MSC of higher passages further enhanced their proliferation rate. Inhibition of mitogen-activated protein kinase 1 (MAPK1) impaired proliferation and differentiation of HPC, but not maintenance of long-term culture initiating cells. siRNA knockdown of Ncadherin and VCAM1 in feeder layer cells increased the fraction of slow dividing HPC, whereas knockdown of integrin beta 1 (ITGB1) and CD44 impaired their differentiation. In conclusion, MSC support proliferation as well as self-renewal of HPC with primitive immunophenotype. The use of early passages of MSC and genetic manipulation of proteins involved in HPC-MSC interaction might further enhance cord blood expansion on MSC. (ProQuest: ... denotes formulae/symbols omitted.)
Mesenchymal stromal cells (MSC) have been suggested to provide a suitable cellular environment for in vitro expansion of haematopoietic stem and progenitor cells (HPC) from umbilical cord blood. In this study, we have simultaneously analysed the cell division history and immunophenotypic differentiation of HPC by using cell division tracking with carboxyfluorescein diacetate N-succinimidyl ester (CFSE). Co-culture with MSC greatly enhanced proliferation of human HPC, especially of the more primitive CD34(+)CD38(-) fraction. Without co-culture CD34 and CD133 expressions decreased after several cell divisions, whereas CD38 expression was up-regulated after some cell divisions and then diminished in fast proliferating cells. Co-culture with MSC maintained a primitive immunophenotype (CD34(+), CD133(+) and CD38(-)) for more population doublings, whereas up-regulation of differentiation markers (CD13, CD45 and CD56) in HPC was delayed to higher numbers of cell divisions. Especially MSC of early cell passages maintained CD34 expression in HPC over more cell divisions, whereas MSC of higher passages further enhanced their proliferation rate. Inhibition of mitogen-activated protein kinase 1 (MAPK1) impaired proliferation and differentiation of HPC, but not maintenance of long-term culture initiating cells. siRNA knockdown of N-cadherin and VCAM1 in feeder layer cells increased the fraction of slow dividing HPC, whereas knockdown of integrin beta 1 (ITGB1) and CD44 impaired their differentiation. In conclusion, MSC support proliferation as well as self-renewal of HPC with primitive immunophenotype. The use of early passages of MSC and genetic manipulation of proteins involved in HPC-MSC interaction might further enhance cord blood expansion on MSC.
Mesenchymal stromal cells (MSC) have been suggested to provide a suitable cellular environment for in vitro expansion of haematopoietic stem and progenitor cells (HPC) from umbilical cord blood. In this study, we have simultaneously analysed the cell division history and immunophenotypic differentiation of HPC by using cell division tracking with carboxyfluorescein diacetate N-succinimidyl ester (CFSE). Co-culture with MSC greatly enhanced proliferation of human HPC, especially of the more primitive CD34(+)CD38(-) fraction. Without co-culture CD34 and CD133 expressions decreased after several cell divisions, whereas CD38 expression was up-regulated after some cell divisions and then diminished in fast proliferating cells. Co-culture with MSC maintained a primitive immunophenotype (CD34(+), CD133(+) and CD38(-)) for more population doublings, whereas up-regulation of differentiation markers (CD13, CD45 and CD56) in HPC was delayed to higher numbers of cell divisions. Especially MSC of early cell passages maintained CD34 expression in HPC over more cell divisions, whereas MSC of higher passages further enhanced their proliferation rate. Inhibition of mitogen-activated protein kinase 1 (MAPK1) impaired proliferation and differentiation of HPC, but not maintenance of long-term culture initiating cells. siRNA knockdown of N-cadherin and VCAM1 in feeder layer cells increased the fraction of slow dividing HPC, whereas knockdown of integrin beta 1 (ITGB1) and CD44 impaired their differentiation. In conclusion, MSC support proliferation as well as self-renewal of HPC with primitive immunophenotype. The use of early passages of MSC and genetic manipulation of proteins involved in HPC-MSC interaction might further enhance cord blood expansion on MSC.Mesenchymal stromal cells (MSC) have been suggested to provide a suitable cellular environment for in vitro expansion of haematopoietic stem and progenitor cells (HPC) from umbilical cord blood. In this study, we have simultaneously analysed the cell division history and immunophenotypic differentiation of HPC by using cell division tracking with carboxyfluorescein diacetate N-succinimidyl ester (CFSE). Co-culture with MSC greatly enhanced proliferation of human HPC, especially of the more primitive CD34(+)CD38(-) fraction. Without co-culture CD34 and CD133 expressions decreased after several cell divisions, whereas CD38 expression was up-regulated after some cell divisions and then diminished in fast proliferating cells. Co-culture with MSC maintained a primitive immunophenotype (CD34(+), CD133(+) and CD38(-)) for more population doublings, whereas up-regulation of differentiation markers (CD13, CD45 and CD56) in HPC was delayed to higher numbers of cell divisions. Especially MSC of early cell passages maintained CD34 expression in HPC over more cell divisions, whereas MSC of higher passages further enhanced their proliferation rate. Inhibition of mitogen-activated protein kinase 1 (MAPK1) impaired proliferation and differentiation of HPC, but not maintenance of long-term culture initiating cells. siRNA knockdown of N-cadherin and VCAM1 in feeder layer cells increased the fraction of slow dividing HPC, whereas knockdown of integrin beta 1 (ITGB1) and CD44 impaired their differentiation. In conclusion, MSC support proliferation as well as self-renewal of HPC with primitive immunophenotype. The use of early passages of MSC and genetic manipulation of proteins involved in HPC-MSC interaction might further enhance cord blood expansion on MSC.
Mesenchymal stromal cells (MSC) have been suggested to provide a suitable cellular environment for in vitro expansion of haematopoietic stem and progenitor cells (HPC) from umbilical cord blood. In this study, we have simultaneously analysed the cell division history and immunophenotypic differentiation of HPC by using cell division tracking with carboxyfluorescein diacetate N-succinimidyl ester (CFSE). Co-culture with MSC greatly enhanced proliferation of human HPC, especially of the more primitive CD34+CD38− fraction. Without co-culture CD34 and CD133 expressions decreased after several cell divisions, whereas CD38 expression was up-regulated after some cell divisions and then diminished in fast proliferating cells. Co-culture with MSC maintained a primitive immunophenotype (CD34+, CD133+ and CD38−) for more population doublings, whereas up-regulation of differentiation markers (CD13, CD45 and CD56) in HPC was delayed to higher numbers of cell divisions. Especially MSC of early cell passages maintained CD34 expression in HPC over more cell divisions, whereas MSC of higher passages further enhanced their proliferation rate. Inhibition of mitogen-activated protein kinase 1 (MAPK1) impaired proliferation and differentiation of HPC, but not maintenance of long-term culture initiating cells. siRNA knockdown of N-cadherin and VCAM1 in feeder layer cells increased the fraction of slow dividing HPC, whereas knockdown of integrin beta 1 (ITGB1) and CD44 impaired their differentiation. In conclusion, MSC support proliferation as well as self-renewal of HPC with primitive immunophenotype. The use of early passages of MSC and genetic manipulation of proteins involved in HPC–MSC interaction might further enhance cord blood expansion on MSC.
Mesenchymal stromal cells (MSC) have been suggested to provide a suitable cellular environment for in vitro expansion of haematopoietic stem and progenitor cells (HPC) from umbilical cord blood. In this study, we have simultaneously analysed the cell division history and immunophenotypic differentiation of HPC by using cell division tracking with carboxyfluorescein diacetate N ‐succinimidyl ester (CFSE). Co‐culture with MSC greatly enhanced proliferation of human HPC, especially of the more primitive CD34 + CD38 − fraction. Without co‐culture CD34 and CD133 expressions decreased after several cell divisions, whereas CD38 expression was up‐regulated after some cell divisions and then diminished in fast proliferating cells. Co‐culture with MSC maintained a primitive immunophenotype (CD34 + , CD133 + and CD38 − ) for more population doublings, whereas up‐regulation of differentiation markers (CD13, CD45 and CD56) in HPC was delayed to higher numbers of cell divisions. Especially MSC of early cell passages maintained CD34 expression in HPC over more cell divisions, whereas MSC of higher passages further enhanced their proliferation rate. Inhibition of mitogen‐activated protein kinase 1 (MAPK1) impaired proliferation and differentiation of HPC, but not maintenance of long‐term culture initiating cells. siRNA knockdown of N‐cadherin and VCAM1 in feeder layer cells increased the fraction of slow dividing HPC, whereas knockdown of integrin beta 1 (ITGB1) and CD44 impaired their differentiation. In conclusion, MSC support proliferation as well as self‐renewal of HPC with primitive immunophenotype. The use of early passages of MSC and genetic manipulation of proteins involved in HPC–MSC interaction might further enhance cord blood expansion on MSC.
Author Eckstein, Volker
Ho, Anthony D.
Saffrich, Rainer
Diehlmann, Anke
Wein, Frederik
Bork, Simone
Walenda, Thomas
Horn, Patrick
Wagner, Wolfgang
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  surname: Horn
  fullname: Horn, Patrick
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  givenname: Frederik
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/19432817$$D View this record in MEDLINE/PubMed
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ContentType Journal Article
Copyright 2009 The Authors Journal compilation © 2010 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd
Copyright Blackwell Publishing Ltd. Jan/Feb 2010
2010. This work is published under https://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.
2009 The Authors Journal compilation © 2010 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd 2009
Copyright_xml – notice: 2009 The Authors Journal compilation © 2010 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd
– notice: Copyright Blackwell Publishing Ltd. Jan/Feb 2010
– notice: 2010. This work is published under https://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.
– notice: 2009 The Authors Journal compilation © 2010 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd 2009
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SSID ssj0036139
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Snippet Mesenchymal stromal cells (MSC) have been suggested to provide a suitable cellular environment for in vitro expansion of haematopoietic stem and progenitor...
Mesenchymal stromal cells (MSC) have been suggested to provide a suitable cellular environment for in vitro expansion of haematopoietic stem and progenitor...
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StartPage 337
SubjectTerms adhesion proteins
ADP-ribosyl Cyclase 1 - metabolism
Antibodies
Antigens, CD34 - metabolism
Biotechnology
Carboxyfluorescein diacetate
CD13 antigen
CD34 antigen
CD38 antigen
CD44 antigen
CD45 antigen
CD56 antigen
Cell Adhesion Molecules - genetics
Cell Adhesion Molecules - metabolism
Cell culture
Cell Culture Techniques
Cell differentiation
Cell Differentiation - physiology
Cell division
Cell Proliferation
Cell self-renewal
Cells, Cultured
Cellular Senescence - physiology
Cloning
Coculture Techniques
Cord blood
co‐culture
Fetal Blood - cytology
Flow cytometry
Genetics
Genotype & phenotype
haematopoietic stem cells
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - physiology
Hemopoiesis
Human subjects
Humans
immunophenotype
Immunophenotyping
Kinases
Laboratories
MAP kinase
Mesenchymal stem cells
mesenchymal stromal cells
Mesenchymal Stromal Cells - cytology
Mesenchymal Stromal Cells - physiology
Mitogen-Activated Protein Kinase 1 - antagonists & inhibitors
Mitogen-Activated Protein Kinase 1 - genetics
Mitogen-Activated Protein Kinase 1 - metabolism
N-Cadherin
Progenitor cells
proliferation
Proteins
replicative senescence
RNA, Small Interfering - genetics
RNA, Small Interfering - metabolism
siRNA
stem cell niche
Stem cells
Stromal cells
Stromal Cells - cytology
Stromal Cells - physiology
Studies
Umbilical cord
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Title Co‐culture with mesenchymal stromal cells increases proliferation and maintenance of haematopoietic progenitor cells
URI https://onlinelibrary.wiley.com/doi/abs/10.1111%2Fj.1582-4934.2009.00776.x
https://www.ncbi.nlm.nih.gov/pubmed/19432817
https://www.proquest.com/docview/217187647
https://www.proquest.com/docview/3075962300
https://www.proquest.com/docview/733591497
https://pubmed.ncbi.nlm.nih.gov/PMC3837622
Volume 14
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