PLoS One

Nilay Chakraborty, Harvard Medical School
Michael A. Menze, Eastern Illinois University
Jason Malsam, University of Minnesota - Twin Cities
Alptekin Aksan, University of Minnesota - Twin Cities
Steven C. Hand, Louisiana State University
Mehmet Toner, Harvard Medical School

This article is available as an open access full text download from http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0024916

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 (LN2) 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.