Telomerase reconstitution immortalizes human fetal hepatocytes without disrupting their differentiation potential
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Telomerase reconstitution immortalizes human fetal hepatocytes without disrupting their differentiation potential. / Wege, Henning; Le, Hai T; Chui, Michael S; Liu, Li; Wu, Jian; Giri, Ranjit; Malhi, Harmeet; Sappal, Baljit S; Kumaran, Vinay; Gupta, Sanjeev; Zern, Mark A.
in: GASTROENTEROLOGY, Jahrgang 124, Nr. 2, 01.02.2003, S. 432-44.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Telomerase reconstitution immortalizes human fetal hepatocytes without disrupting their differentiation potential
AU - Wege, Henning
AU - Le, Hai T
AU - Chui, Michael S
AU - Liu, Li
AU - Wu, Jian
AU - Giri, Ranjit
AU - Malhi, Harmeet
AU - Sappal, Baljit S
AU - Kumaran, Vinay
AU - Gupta, Sanjeev
AU - Zern, Mark A
PY - 2003/2/1
Y1 - 2003/2/1
N2 - BACKGROUND & AIMS: The availability of in vitro expandable human hepatocytes would greatly advance liver-directed cell therapies. Therefore, we examined whether human fetal hepatocytes are amenable to telomerase-mediated immortalization without inducing a transformed phenotype and disrupting their differentiation potential. Telomerase is a ribonucleoprotein that plays a pivotal role in maintaining telomere length and chromosome stability. Human somatic cells, including hepatocytes, exhibit no telomerase activity. Consequently, their telomeres progressively shorten with each cell cycle until critically short telomeres trigger replicative senescence.METHODS: The catalytic subunit, telomerase reverse transcriptase, was expressed in human fetal hepatocytes. Transduced cells were characterized for telomerase activity, telomere length, proliferative capacity, hepatocellular functions, oncogenicity, and their in vivo maturation potential.RESULTS: The expression of human telomerase reverse transcriptase restored telomerase activity in human fetal hepatocytes. Telomerase-reconstituted cells were capable of preserving elongated telomeres, propagated in culture beyond replicative senescence for more than 300 cell doublings (to date), and maintained their liver-specific nature, as analyzed by a panel of hepatic growth factors, growth factor receptors, and transcription factors as well as albumin, glucose-6-phosphatase, glycogen synthesis, cytochrome P450 (CYP) expression profiles, and urea production. Moreover, the immortalized cells exhibited no oncogenicity, and no up-regulation of c-Myc was detected. The cells engrafted and survived in the liver of immunodeficient mice with hepatocellular gene expression.CONCLUSIONS: Reconstitution of telomerase activity induces indefinite replication in human fetal hepatocytes and offers unique opportunities for examining basic biologic mechanisms and for considering development of stable cell lines for liver-directed therapies.
AB - BACKGROUND & AIMS: The availability of in vitro expandable human hepatocytes would greatly advance liver-directed cell therapies. Therefore, we examined whether human fetal hepatocytes are amenable to telomerase-mediated immortalization without inducing a transformed phenotype and disrupting their differentiation potential. Telomerase is a ribonucleoprotein that plays a pivotal role in maintaining telomere length and chromosome stability. Human somatic cells, including hepatocytes, exhibit no telomerase activity. Consequently, their telomeres progressively shorten with each cell cycle until critically short telomeres trigger replicative senescence.METHODS: The catalytic subunit, telomerase reverse transcriptase, was expressed in human fetal hepatocytes. Transduced cells were characterized for telomerase activity, telomere length, proliferative capacity, hepatocellular functions, oncogenicity, and their in vivo maturation potential.RESULTS: The expression of human telomerase reverse transcriptase restored telomerase activity in human fetal hepatocytes. Telomerase-reconstituted cells were capable of preserving elongated telomeres, propagated in culture beyond replicative senescence for more than 300 cell doublings (to date), and maintained their liver-specific nature, as analyzed by a panel of hepatic growth factors, growth factor receptors, and transcription factors as well as albumin, glucose-6-phosphatase, glycogen synthesis, cytochrome P450 (CYP) expression profiles, and urea production. Moreover, the immortalized cells exhibited no oncogenicity, and no up-regulation of c-Myc was detected. The cells engrafted and survived in the liver of immunodeficient mice with hepatocellular gene expression.CONCLUSIONS: Reconstitution of telomerase activity induces indefinite replication in human fetal hepatocytes and offers unique opportunities for examining basic biologic mechanisms and for considering development of stable cell lines for liver-directed therapies.
KW - Animals
KW - Cell Differentiation
KW - Cell Line, Transformed
KW - DNA-Binding Proteins
KW - Female
KW - Fetus
KW - Hepatocytes
KW - Humans
KW - Mice
KW - Neoplasms
KW - Pregnancy
KW - Telomerase
KW - Telomere
KW - Transduction, Genetic
U2 - 10.1053/gast.2003.50064
DO - 10.1053/gast.2003.50064
M3 - SCORING: Journal article
C2 - 12557149
VL - 124
SP - 432
EP - 444
JO - GASTROENTEROLOGY
JF - GASTROENTEROLOGY
SN - 0016-5085
IS - 2
ER -