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Related Experiment Video

Updated: May 12, 2026

Bulk Droplet Vitrification for Primary Hepatocyte Preservation
11:07

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Published on: October 25, 2019

Forced-convective vitrification with liquid cryogens.

Shaw-Ruey Lyu1, Jen-Hung Huang, Wei-Hung Shih

  • 1Joint Center, Tzu-Chi Dalin General Hospital, Chia-Yi and Tzu-Chi University, Hua-Lien, Taiwan, ROC.

Cryobiology
|April 3, 2013
PubMed
Summary

Forced-convective vitrification using sub-boiling liquid oxygen (LOX(bbp)) significantly enhances cooling rates. This innovative method reduces cryoprotective agent (CPA) concentrations needed for vitrification, minimizing toxicity and osmotic stress during cell preservation.

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Area of Science:

  • Cryobiology
  • Biotechnology
  • Materials Science

Background:

  • Cell cryopreservation via vitrification typically demands high, toxic cryoprotectant agent (CPA) concentrations.
  • High CPA concentrations lead to detrimental osmotic stresses during addition and removal.

Purpose of the Study:

  • To develop an innovative cell cryopreservation technique to reduce CPA concentration.
  • To enhance cooling rates during vitrification using forced convection and novel cryogens.

Main Methods:

  • Proposed forced-convective vitrification using sub-boiling liquid oxygen (LOX(bbp)) as the cryogen.
  • Applied constant sample velocity to mitigate insulating gas bubbles.
  • Varied cryogen type, sample velocity, and test solution volume.

Main Results:

  • LOX(bbp) achieved a 2.3-fold greater cooling rate than liquid nitrogen (LN(abp)) at 1.2 m/s.
  • Increasing sample velocity from 0.2 to 1.2 m/s with LOX(bbp) enhanced cooling rate by 1.9 times.
  • Reduced CPA concentration to 25% required for vitrification using LOX(bbp) at 1.2 m/s and 1.73 μl solution volume.

Conclusions:

  • Forced-convective vitrification with LOX(bbp) effectively increases cooling rates.
  • This method significantly lowers the required CPA concentration for successful vitrification.
  • The technique minimizes CPA-related toxicity and osmotic stress in cell cryopreservation.