title: Supercooling as a viable non-freezing cell preservation method of rat hepatocytes.
abstract: Supercooling preservation holds the potential to drastically extend the preservation time of organs, tissues and engineered tissue products, and fragile cell types that do not lend themselves well to cryopreservation or vitrification. Here, we investigate the effects of supercooling preservation (SCP at -4(o)C) on primary rat hepatocytes stored in cryovials and compare its success (high viability and good functional characteristics) to that of static cold storage (CS at +4(o)C) and cryopreservation. We consider two prominent preservation solutions a) Hypothermosol (HTS-FRS) and b) University of Wisconsin solution (UW) and a range of preservation temperatures (-4 to -10 (o)C). We find that there exists an optimum temperature (-4(o)C) for SCP of rat hepatocytes which yields the highest viability; at this temperature HTS-FRS significantly outperforms UW solution in terms of viability and functional characteristics (secretions and enzymatic activity in suspension and plate culture). With the HTS-FRS solution we show that the cells can be stored for up to a week with high viability (~56%); moreover we also show that the preservation can be performed in large batches (50 million cells) with equal or better viability and no loss of functionality as compared to smaller batches (1.5 million cells) performed in cryovials.
authors.affiliation: Center for Engineering in Medicine at Massachusetts General Hospital, Harvard Medical School and Shriners Hospital for Children, Boston, Massachusetts, United States of America.
authors.forename: O Berk
authors.initials: OB
authors.lastname: Usta
authors.forename: Yeonhee
authors.initials: Y
authors.lastname: Kim
authors.forename: Sinan
authors.initials: S
authors.lastname: Ozer
authors.forename: Bote G
authors.initials: BG
authors.lastname: Bruinsma
authors.forename: Jungwoo
authors.initials: J
authors.lastname: Lee
authors.forename: Esin
authors.initials: E
authors.lastname: Demir
authors.forename: Tim A
authors.initials: TA
authors.lastname: Berendsen
authors.forename: Catheleyne F
authors.initials: CF
authors.lastname: Puts
authors.forename: Maria-Louisa
authors.initials: ML
authors.lastname: Izamis
authors.forename: Korkut
authors.initials: K
authors.lastname: Uygun
authors.forename: Basak E
authors.initials: BE
authors.lastname: Uygun
authors.forename: Martin L
authors.initials: ML
authors.lastname: Yarmush
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D022781:Hepatocytes / Q000166:cytology*; D019188:Organ Preservation Solutions; D051381:Rats; D012996:Solutions; D013696:Temperature
chemical_list: D019188:Organ Preservation Solutions; D012996:Solutions