Human short-term repopulating cells have enhanced telomerase reverse transcriptase expression.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/155994
- author
- Järås, Marcus LU ; Edqvist, Anna LU ; Rebetz, Johan LU ; Salford, Leif LU ; Widegren, Bengt LU and Fan, Xiaolong LU
- organization
- publishing date
- 2006
- 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
- 2016-04-01 12:34:38
- date last changed
- 2022-04-13 20:55:32
@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 < .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.}}, 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}}, doi = {{10.1182/blood-2005-09-008904}}, volume = {{108}}, year = {{2006}}, }