Induction of telomere shortening and replicative senescence by cryopreservation.
Cryopreservation can alter cellular function under certain conditions. In this report, we demonstrate the induction of cellular senescence after cells have been cryopreserved using a standard protocol. A retinal pigment epithelial cell line frozen at a specific freezing rate and subsequently thawed showed severely impaired proliferation compared to cells that were not cryopreserved. The induction of senescence was suggested by senescent associated beta-galactosidase activity and diminished bromo-deoxyuridine incorporation. A remarkable increase of single-strand DNA breaks in terminal restriction fragment (TRF) were found in cryopreserved cells immediately after thawing. The rate of mean TRF length shortening was accelerated after cryopreservation. Given this evidence, we hypothesize that cryopreservation may cause telomere shortening and cellular senescence under certain freezing conditions.
Honda
S
S
Department of Ophthalmology, University of California, Davis 95616-8794, USA.
Weigel
A
A
Hjelmeland
L M
LM
Handa
J T
JT
D001973:Bromodeoxyuridine / Q000378:metabolism; D002455:Cell Division; D002460:Cell Line; D016922:Cellular Senescence; D015925:Cryopreservation; D004249:DNA Damage; D006801:Humans; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015719:Single-Strand Specific DNA and RNA Endonucleases; D016615:Telomere / Q000378:metabolism*; D001616:beta-Galactosidase / Q000378:metabolism
D017382:Reactive Oxygen Species; D015719:Single-Strand Specific DNA and RNA Endonucleases; D001616:beta-Galactosidase; D001973:Bromodeoxyuridine