title: Intracellular trehalose improves the survival of cryopreserved mammalian cells.
abstract: We report that the introduction of low concentrations of intracellular trehalose can greatly improve the survival of mammalian cells during cryopreservation. Using a genetically engineered mutant of Staphylococcus aureus alpha-hemolysin to create pores in the cellular membrane, we were able to load trehalose into cells. Low concentrations (0.2 M) of trehalose permitted long-term post-thaw survival of more than 80% of 3T3 fibroblasts and 70% of human keratinocytes. These results indicate that simplified and widely applicable freezing protocols may be possible using sugars as intracellular cryoprotective additives.
authors.affiliation: The Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, MA, 02114, USA.
authors.forename: A
authors.initials: A
authors.lastname: Eroglu
authors.forename: M J
authors.initials: MJ
authors.lastname: Russo
authors.forename: R
authors.initials: R
authors.lastname: Bieganski
authors.forename: A
authors.initials: A
authors.lastname: Fowler
authors.forename: S
authors.initials: S
authors.lastname: Cheley
authors.forename: H
authors.initials: H
authors.lastname: Bayley
authors.forename: M
authors.initials: M
authors.lastname: Toner
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics; D001692:Biological Transport; D002462:Cell Membrane; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D005818:Genetic Engineering; D006460:Hemolysin Proteins / Q000235:genetics; D006801:Humans; D015603:Keratinocytes; D051379:Mice; D013211:Staphylococcus aureus / Q000235:genetics; D014199:Trehalose / Q000494:pharmacology*
chemical_list: D001427:Bacterial Toxins; D003451:Cryoprotective Agents; D006460:Hemolysin Proteins; C021091:staphylococcal alpha-toxin; D014199:Trehalose