title: Cryopreservation of spin-dried mammalian cells.
abstract: This study reports an alternative approach to achieve vitrification where cells are pre-desiccated prior to cooling to cryogenic temperatures for storage. Chinese Hamster Ovary (CHO) cells suspended in a trehalose solution were rapidly and uniformly desiccated to a low moisture content (<0.12 g of water per g of dry weight) using a spin-drying technique. Trehalose was also introduced into the cells using a high-capacity trehalose transporter (TRET1). Fourier Transform Infrared Spectroscopy (FTIR) was used to examine the uniformity of water concentration distribution in the spin-dried samples. 62% of the cells were shown to survive spin-drying in the presence of trehalose following immediate rehydration. The spin-dried samples were stored in liquid nitrogen (LN(2)) at a vitrified state. It was shown that following re-warming to room temperature and re-hydration with a fully complemented cell culture medium, 51% of the spin-dried and vitrified cells survived and demonstrated normal growth characteristics. Spin-drying is a novel strategy that can be used to improve cryopreservation outcome by promoting rapid vitrification.
authors.affiliation: Center for Engineering in Medicine and BioMEMS Resource Center, Massachusetts General Hospital, Harvard Medical School and Shriners Hospital for Children, Boston, Massachusetts, United States of America. chakraborty.nilay@mgh.harvard.edu
authors.forename: Nilay
authors.initials: N
authors.lastname: Chakraborty
authors.forename: Michael A
authors.initials: MA
authors.lastname: Menze
authors.forename: Jason
authors.initials: J
authors.lastname: Malsam
authors.forename: Alptekin
authors.initials: A
authors.lastname: Aksan
authors.forename: Steven C
authors.initials: SC
authors.lastname: Hand
authors.forename: Mehmet
authors.initials: M
authors.lastname: Toner
mesh_terms: D000818:Animals; D016466:CHO Cells; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured / Q000166:cytology*; D006224:Cricetinae; D015925:Cryopreservation / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D009584:Nitrogen / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods; D013696:Temperature; D014199:Trehalose / Q000737:chemistry; D058989:Vitrification; D014867:Water / Q000737:chemistry
chemical_list: D003470:Culture Media; D011108:Polymers; D014867:Water; D014199:Trehalose; D009584:Nitrogen