Cryopreservation of spin-dried mammalian cells

Nilay Chakraborty, Michael A. Menze, Jason Malsam, Alptekin Aksan, Steven C. Hand, Mehmet Toner

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9 Scopus citations

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.

Original languageEnglish (US)
Article numbere24916
JournalPloS one
Volume6
Issue number9
DOIs
StatePublished - Sep 22 2011

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    Chakraborty, N., Menze, M. A., Malsam, J., Aksan, A., Hand, S. C., & Toner, M. (2011). Cryopreservation of spin-dried mammalian cells. PloS one, 6(9), [e24916]. https://doi.org/10.1371/journal.pone.0024916