Cryopreservation of spin-dried mammalian cells

PLoS One. 2011;6(9):e24916. doi: 10.1371/journal.pone.0024916. Epub 2011 Sep 22.

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • CHO Cells
  • Cell Culture Techniques*
  • Cell Survival
  • Cells, Cultured / cytology*
  • Cricetinae
  • Cryopreservation / methods*
  • Culture Media / chemistry
  • Nitrogen / chemistry
  • Polymers / chemistry
  • Spectroscopy, Fourier Transform Infrared / methods
  • Temperature
  • Trehalose / chemistry
  • Vitrification
  • Water / chemistry

Substances

  • Culture Media
  • Polymers
  • Water
  • Trehalose
  • Nitrogen