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Bulk Droplet Vitrification for Primary Hepatocyte Preservation
Published on: October 25, 2019
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Bulk Droplet Vitrification for Primary Hepatocyte Preservation.
Reinier J de Vries1, Peony D Banik2, Sonal Nagpal2
1Center for Engineering in Medicine, Harvard Medical School, Massachusetts General Hospital; Shriners Hospitals for Children, Boston; Department of Surgery, University of Amsterdam.
Journal of Visualized Experiments : Jove
|November 12, 2019
Summary
Bulk droplet vitrification offers ice-free cryopreservation for larger samples. This method minimizes cryoprotective agent toxicity, improving cell viability and function compared to traditional slow-freezing.
Area of Science:
- Cryobiology
- Biotechnology
- Cell Biology
Background:
- Vitrification is an ice-free cryopreservation technique.
- High cooling rates are essential to prevent ice crystal formation.
- Conventional vitrification requires high cryoprotective agent (CPA) concentrations, leading to toxicity and sample size limitations.
Purpose of the Study:
- To detail the methodology of bulk droplet vitrification.
- To present a novel approach overcoming limitations of traditional vitrification.
- To demonstrate improved cryopreservation of biological samples.
Main Methods:
- Cells are pre-incubated with low intracellular CPA concentrations.
- Rapid osmotic dehydration concentrates CPA before vitrification.
- A fluidic device generates large droplets for vitrification in liquid nitrogen.
Main Results:
- Bulk droplet vitrification enables ice-free cryopreservation of larger sample volumes.
- The method significantly reduces CPA toxicity.
- Increased hepatocyte viability and metabolic function were observed compared to slow-freezing.
Conclusions:
- Bulk droplet vitrification is a viable alternative to slow-freezing.
- This technique minimizes CPA toxicity and is suitable for large cell quantities.
- Improved cell viability and function are key advantages for cryopreservation.
