Isothermal vitrification methodology development for non-cryogenic storage of archival human sera

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Biorepositories worldwide collect human serum samples and store them for future research. Currently, hundreds of biorepositories across the world store human serum samples in refrigerators, freezers, or liquid nitrogen without following any specific cryopreservation protocol. This method of storage is both expensive and potentially detrimental to the biospecimens. To decrease both cost of storage and the freeze/thaw stresses, we explored the feasibility of storing archival human serum samples at non-cryogenic temperatures using isothermal vitrification. When biospecimens are vitrified, biochemical reactions can be stopped, the specimen ceases to degrade, and macromolecules can be stabilized without requiring cryogenic storage. In this study, 0.2, 0.4, or 0.8M trehalose; 0, 0.005 or 0.01M dextran; and 0 or 10% (v/v) glycerol was added to human serum samples. The samples were either dried diffusively as sessile droplets or desiccated under vacuum after they are adsorbed onto glass microfiber filters. The glass transition temperatures (Tg) of the desiccated samples were measured by temperature-ramp Fourier Transform Infrared (FTIR) spectroscopy. Sera samples vitrified at 4±2°C when 0.8M trehalose and 0.01M dextran were added and the samples were vacuum dried for two hours. Western immunoblotting showed that vitrified serum proteins were minimally degraded when stored for up to one month at 4°C. About 80% of all proteins were recovered after storage at 4°C on glass microfiber filters, and recovery did not decrease with storage time. These results demonstrated the feasibility of long-term storage of vitrified serum at hypothermic (and non-cryogenic) temperatures.

Original languageEnglish (US)
Pages (from-to)176-185
Number of pages10
JournalCryobiology
Volume66
Issue number2
DOIs
StatePublished - Apr 1 2013

Fingerprint

Vitrification
vitrification
Trehalose
Dextrans
Serum
Vacuum
Glass
Refrigerators
Liquid nitrogen
Macromolecules
sampling
Cryogenics
Glycerol
Temperature
Blood Proteins
dextran
trehalose
methodology
Recovery
glass

Keywords

  • Desiccation
  • Glass transition temperature
  • Isothermal vitrification
  • Non-cryogenic storage
  • Serum
  • Stabilization
  • Trehalose

Cite this

Isothermal vitrification methodology development for non-cryogenic storage of archival human sera. / Less, Rebekah; Boylan, Kristin L; Skubitz, Amy P; Aksan, Alptekin.

In: Cryobiology, Vol. 66, No. 2, 01.04.2013, p. 176-185.

Research output: Contribution to journalArticle

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