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Bulk Droplet Vitrification for Primary Hepatocyte Preservation
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Accelerating cryoprotectant delivery using vacuum infiltration.

Ryan J Forcier1, Robert T Heussner1, Lauren Newsom2

  • 1School of Chemical, Biological and Environmental Engineering, Oregon State University, Corvallis, OR, USA.

Cryobiology
|July 14, 2023
PubMed
Summary
This summary is machine-generated.

Cryopreservation of vertebral bodies (VBs) is difficult. Carbonation and vacuum cycling may slightly improve cryoprotectant delivery into large tissues like VBs.

Keywords:
CryoprotectantMass transferOrganTissueVacuumVertebral body

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Area of Science:

  • Biomedical Engineering
  • Tissue Engineering
  • Cryobiology

Background:

  • Cryopreservation of large biological structures, such as vertebral bodies (VBs), is limited by mass transport challenges.
  • Effective delivery of cryoprotective agents into dense tissues like VBs is crucial for successful cryopreservation.
  • Current methods struggle to achieve uniform cryoprotectant distribution in large tissue volumes.

Purpose of the Study:

  • To investigate the efficacy of carbonation and vacuum cycling for enhancing cryoprotectant penetration into VBs.
  • To determine if CO2 gas infiltration followed by vacuum cycling can improve cryoprotectant delivery in large tissue structures.
  • To assess the potential of this method for advancing VB cryopreservation for clinical and research applications.

Main Methods:

  • Utilized agarose gel and colored dye as a model system to evaluate penetration enhancement.
  • Applied carbonation (high-pressure CO2 gas) followed by vacuum cycling to the model system and human VBs.
  • Employed CT imaging to visualize gas bubbles and assess DMSO concentration within VBs.

Main Results:

  • Carbonation and vacuum cycling increased dye penetration by 14% in the agarose gel model.
  • CT imaging confirmed the presence of gas bubbles in VBs, with vacuum cycling reducing bubble volume by over 50%.
  • A statistically significant increase in DMSO concentration within VBs was not detected via CT imaging.

Conclusions:

  • Carbonation and vacuum cycling show potential for modest improvement in cryoprotectant delivery into large tissue structures like VBs.
  • Further research is needed to optimize the technique and confirm its effectiveness for VB cryopreservation.
  • This method could offer benefits for preserving large tissues for transplantation or research purposes.