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Human short-term repopulating cells have enhanced telomerase reverse transcriptase expression.

Järås, Marcus LU ; Edqvist, Anna LU ; Rebetz, Johan LU ; Salford, Leif LU ; Widegren, Bengt LU and Fan, Xiaolong LU (2006) In Blood 108(3). p.1084-1091
Abstract
Telomerase activity has been suggested to be critically involved in hematopoietic stem cell (HSC) self-renewal. However, it has been unclear whether human HSCs have telomerase activity and how telomerase activity is regulated within the HSC and progenitor pool. Here, we isolated living cord-blood (CB) CD34+ cells with up-regulated human telomerase reverse transcriptase (hTERT) expression by using an hTERT-reporting adenoviral vector encoding destabilized green fluorescent protein (dGFP) driven by the hTERT promoter, and functionally characterized them in comparison with control vector–transduced CD34+ cells expressing GFP. Following a 2-day serum-free transduction protocol, cells were sorted into a dGFP+ and a GFP+ fraction. Cell-cycle... (More)
Telomerase activity has been suggested to be critically involved in hematopoietic stem cell (HSC) self-renewal. However, it has been unclear whether human HSCs have telomerase activity and how telomerase activity is regulated within the HSC and progenitor pool. Here, we isolated living cord-blood (CB) CD34+ cells with up-regulated human telomerase reverse transcriptase (hTERT) expression by using an hTERT-reporting adenoviral vector encoding destabilized green fluorescent protein (dGFP) driven by the hTERT promoter, and functionally characterized them in comparison with control vector–transduced CD34+ cells expressing GFP. Following a 2-day serum-free transduction protocol, cells were sorted into a dGFP+ and a GFP+ fraction. Cell-cycle analysis revealed that the dGFP+ cells had a greater proportion of cells in S/G2/M phase compared with the GFP+ cells, (56% ± 1.8% vs 35% ± 4.3%; P < .001) and fewer cells in G0 phase (8.1% ± 3.0% vs 20% ± 4.7%; P < .01) However, the colony-forming and short-term nonobese diabetic/severe combined immunodeficient (NOD/SCID) B2m–/– mice bone marrow–repopulating capacities were similar between the dGFP+ and the GFP+ cells. Interestingly, the dGFP+ cells had a 6-fold lower repopulating capacity in NOD/SCID mice compared with the GFP+ cells and lacked secondary NOD/SCID B2m–/– mice bone marrow–repopulating capacity. Thus, up-regulation of hTERT expression within the CB HSC pool is accompanied by decreased self-renewal capacity. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Blood
volume
108
issue
3
pages
1084 - 1091
publisher
American Society of Hematology
external identifiers
  • pmid:16861355
  • wos:000239381000052
  • scopus:33746640430
ISSN
1528-0020
DOI
10.1182/blood-2005-09-008904
language
English
LU publication?
yes
id
e617cc8f-db62-45d2-8cc8-11600fe131b0 (old id 155994)
date added to LUP
2007-07-11 12:00:40
date last changed
2019-03-05 02:04:35
@article{e617cc8f-db62-45d2-8cc8-11600fe131b0,
  abstract     = {Telomerase activity has been suggested to be critically involved in hematopoietic stem cell (HSC) self-renewal. However, it has been unclear whether human HSCs have telomerase activity and how telomerase activity is regulated within the HSC and progenitor pool. Here, we isolated living cord-blood (CB) CD34+ cells with up-regulated human telomerase reverse transcriptase (hTERT) expression by using an hTERT-reporting adenoviral vector encoding destabilized green fluorescent protein (dGFP) driven by the hTERT promoter, and functionally characterized them in comparison with control vector–transduced CD34+ cells expressing GFP. Following a 2-day serum-free transduction protocol, cells were sorted into a dGFP+ and a GFP+ fraction. Cell-cycle analysis revealed that the dGFP+ cells had a greater proportion of cells in S/G2/M phase compared with the GFP+ cells, (56% ± 1.8% vs 35% ± 4.3%; P &lt; .001) and fewer cells in G0 phase (8.1% ± 3.0% vs 20% ± 4.7%; P &lt; .01) However, the colony-forming and short-term nonobese diabetic/severe combined immunodeficient (NOD/SCID) B2m–/– mice bone marrow–repopulating capacities were similar between the dGFP+ and the GFP+ cells. Interestingly, the dGFP+ cells had a 6-fold lower repopulating capacity in NOD/SCID mice compared with the GFP+ cells and lacked secondary NOD/SCID B2m–/– mice bone marrow–repopulating capacity. Thus, up-regulation of hTERT expression within the CB HSC pool is accompanied by decreased self-renewal capacity.},
  author       = {Järås, Marcus and Edqvist, Anna and Rebetz, Johan and Salford, Leif and Widegren, Bengt and Fan, Xiaolong},
  issn         = {1528-0020},
  language     = {eng},
  number       = {3},
  pages        = {1084--1091},
  publisher    = {American Society of Hematology},
  series       = {Blood},
  title        = {Human short-term repopulating cells have enhanced telomerase reverse transcriptase expression.},
  url          = {http://dx.doi.org/10.1182/blood-2005-09-008904},
  volume       = {108},
  year         = {2006},
}