Elevated telomerase activity and minimal telomere loss in cord blood long-term cultures with extensive stem cell replication
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Elevated telomerase activity and minimal telomere loss in cord blood long-term cultures with extensive stem cell replication. / Gammaitoni, Loretta; Weisel, Katja C; Gunetti, Monica; Wu, Kai-Da; Bruno, Stefania; Pinelli, Silvana; Bonati, Antonio; Aglietta, Massimo; Moore, Malcolm A S; Piacibello, Wanda.
In: BLOOD, Vol. 103, No. 12, 15.06.2004, p. 4440-8.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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T1 - Elevated telomerase activity and minimal telomere loss in cord blood long-term cultures with extensive stem cell replication
AU - Gammaitoni, Loretta
AU - Weisel, Katja C
AU - Gunetti, Monica
AU - Wu, Kai-Da
AU - Bruno, Stefania
AU - Pinelli, Silvana
AU - Bonati, Antonio
AU - Aglietta, Massimo
AU - Moore, Malcolm A S
AU - Piacibello, Wanda
PY - 2004/6/15
Y1 - 2004/6/15
N2 - Telomerase activity, telomere length, stem/progenitor cell production, and function of CD34+ cells from cord blood (CB), bone marrow, and mobilized peripheral blood were evaluated in long-term cultures. CB cells were cultured either on OP-9 stromal cells transduced with an adenovector expressing thrombopoietin (TPO) or stimulated by a cytokine cocktail in the absence of stroma, with, in one method, CD34+ cells reisolated at monthly intervals for passage. Continuous expansion of stem cells as measured by in vitro cobblestone area and secondary colony-forming assays was noted for 18 to 20 weeks and by severe combined immunodeficiency (SCID)-repopulating cells (SRCs), capable of repopulating and serially passage in nonobese diabetic/SCID mice, for 16 weeks. Despite this extensive proliferation, telomere length initially increased and only at late stages of culture was evidence of telomere shortening noted. This telomere stabilization correlated with maintenance of high levels of telomerase activity in the CD34+ cell population for prolonged periods of culture. Cytokine-stimulated cultures of adult CD34+ cells showed CD34+ and SRC expansion (6-fold) for only 3 to 4 weeks with telomere shortening and low levels of telomerase. There is clearly a clinical value for a system that provides extensive stem cell expansion without concomitant telomere erosion.
AB - Telomerase activity, telomere length, stem/progenitor cell production, and function of CD34+ cells from cord blood (CB), bone marrow, and mobilized peripheral blood were evaluated in long-term cultures. CB cells were cultured either on OP-9 stromal cells transduced with an adenovector expressing thrombopoietin (TPO) or stimulated by a cytokine cocktail in the absence of stroma, with, in one method, CD34+ cells reisolated at monthly intervals for passage. Continuous expansion of stem cells as measured by in vitro cobblestone area and secondary colony-forming assays was noted for 18 to 20 weeks and by severe combined immunodeficiency (SCID)-repopulating cells (SRCs), capable of repopulating and serially passage in nonobese diabetic/SCID mice, for 16 weeks. Despite this extensive proliferation, telomere length initially increased and only at late stages of culture was evidence of telomere shortening noted. This telomere stabilization correlated with maintenance of high levels of telomerase activity in the CD34+ cell population for prolonged periods of culture. Cytokine-stimulated cultures of adult CD34+ cells showed CD34+ and SRC expansion (6-fold) for only 3 to 4 weeks with telomere shortening and low levels of telomerase. There is clearly a clinical value for a system that provides extensive stem cell expansion without concomitant telomere erosion.
KW - Adenoviridae
KW - Animals
KW - Antigens, CD
KW - Antigens, CD34
KW - Cell Division
KW - Fetal Blood
KW - Genetic Vectors
KW - Hematopoietic Stem Cells
KW - Humans
KW - Infant, Newborn
KW - Mice
KW - Mice, Inbred NOD
KW - Mice, SCID
KW - Telomerase
KW - Thrombopoietin
KW - Transfection
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
KW - Research Support, U.S. Gov't, P.H.S.
U2 - 10.1182/blood-2003-09-3079
DO - 10.1182/blood-2003-09-3079
M3 - SCORING: Journal article
C2 - 14726371
VL - 103
SP - 4440
EP - 4448
JO - BLOOD
JF - BLOOD
SN - 0006-4971
IS - 12
ER -