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Related Experiment Video

Updated: Sep 27, 2025

Combination of Microstereolithography and Electrospinning to Produce Membranes Equipped with Niches for Corneal Regeneration
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Development of a Vitrification Preservation Process for Bioengineered Epithelial Constructs.

Lia H Campbell1, Kelvin G M Brockbank1

  • 1Tissue Testing Technologies LLC, 2231 Technical Parkway, Suite A, North Charleston, SC 29406, USA.

Cells
|April 12, 2022
PubMed
Summary

Developing ice-free vitrification methods for human bioengineered epithelial constructs enables cryopreservation and banking, reducing costs and improving delivery times for tissue engineering applications.

Keywords:
bioengineered tissue constructscryopreservationvitrification

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Area of Science:

  • Tissue Engineering
  • Cryobiology
  • Biotechnology

Background:

  • Growing demand for human bioengineered tissue constructs due to reduced animal testing.
  • Current just-in-time manufacturing leads to delays and high costs.
  • Cryopreservation and banking offer solutions for faster delivery and cost reduction.

Purpose of the Study:

  • Develop ice-free vitrification formulations and protocols for human bioengineered epithelial constructs.
  • Scale up protocols from individual constructs to 24-well plates.
  • Ensure long-term viability of cryopreserved tissue constructs.

Main Methods:

  • Vitrification using 55% (VS55) and 70% (VS70) cryoprotectant (CPA) formulations.
  • Optimization of protocols for improved post-rewarming viability.
  • Testing viability in deep-well plates (up to 24 constructs) and storage below -135 °C.

Main Results:

  • Initial single construct viability >80% post-rewarming.
  • Protocol improvements led to sustained viability (>70%) for 1-3 days post-rewarming in 24-well plates.
  • Viability maintained after 2-7 months of storage below -135 °C.

Conclusions:

  • Successful development of ice-free vitrification protocols for human bioengineered epithelial constructs.
  • Demonstrated scalability to 24-well plates with high post-warming and in vitro viability.
  • Established feasibility of long-term cryopreservation for tissue banking and distribution.