Cryopreservation of spin-dried mammalian cells.

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PloS one, ISSN: 1932-6203, Vol: 6, Issue: 9, Page: e24916

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10.1371/journal.pone.0024916; 10.1371/journal.pone.0024916.g001; 10.1371/journal.pone.0024916.g002; 10.1371/journal.pone.0024916.g004; 10.1371/journal.pone.0024916.g005; 10.1371/journal.pone.0024916.g007; 10.1371/journal.pone.0024916.g006; 10.1371/journal.pone.0024916.g003
PMC3178566; 3178566
Nilay Chakraborty; Michael A. Menze; Jason Malsam; Alptekin Aksan; Steven C. Hand; Mehmet Toner; Boris Rubinsky
Public Library of Science (PLoS); Figshare
Medicine; Biochemistry, Genetics and Molecular Biology; Agricultural and Biological Sciences; Cryopreservation; Mammalian Cells; Spin-Dried; Spin-Dry; Cryopreservation; Mammalian Cells; Spin-Dried; Spin-Dry; Physics; Cell Biology; Physiology; Biotechnology; intracellular; trehalose; wild-type; cho; cells; cho-tret1; Biology; spin-drying; drying; configuration; spinning; 10; mm; 20; 120; choline; ph; ftir; spin-dried; solutions; stored; buffer; buffers; rehydrated
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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.