Drug discovery pipelines rely on the availability of isolated primary hepatocytes for investigating potential hepatotoxicity prior to clinical application. These hepatocytes are typically isolated from livers rejected for transplantation and subsequently cryopreserved for later usage. The gold-standard cryopreservation technique, slow-freezing, is a labor-intensive process, with significant post-storage viability loss. In this work, we introduce parallelized droplet vitrification, a technique for high-volumetric, rapid vitrification of suspended cells. We show the utility of this technique through the single-run vitrification of the whole-rate liver hepatocyte yield, resulting in a 1600% increase in single-batch vitrification and a 500% increase in droplet generation rate compared to previous droplet vitrification approaches. Additionally, we showed that these implementations maintained improved post-preservation outcomes in primary rat hepatocytes.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11275928 | PMC |
| http://dx.doi.org/10.1101/2024.07.14.603471 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
An efficient droplet-vitrification cryopreservation procedure for high imperatorin-yielding hairy root clones of Urena lobata.
Cryobiology
December 2024
Plant Biotechnology Laboratory, Department of Plant Biotechnology and Biotransformation, Faculty of Biology and Biotechnology, University of Science, Ho Chi Minh City, Viet Nam; Vietnam National University Ho Chi Minh City, Viet Nam. Electronic address:
Article Synopsis
- Researchers found that imperatorin, a valuable anti-cancer and anti-inflammatory compound, can be extracted from the hairy roots of Urena lobata, but cryo-injury and cryoprotectant toxicity pose challenges for preserving root clones effectively.
- They identified key factors that reduce plasmolysis during cryopreservation and improved a droplet-vitrification technique that included specific concentrations of sucrose and glycerol to enhance root survival rates.
- The new cryopreservation method led to a 93.3% regeneration rate and comparable growth and imperatorin production to untreated roots after multiple subcultures, emphasizing the effectiveness of their plasmolysis evaluation approach for optimizing preservation techniques.
Numerical and experimental investigation of a droplet vitrification process: A thermofluidic analysis for cryopreservation.
Cryobiology
December 2024
HemoCord - Umbilical Cord Blood Bank, Unisinos Technology Park, São Leopoldo, RS, Brazil.
One of the biggest challenges in studying vitrification protocols for small volumes of biological materials, especially the microdroplet vitrification protocol, is measuring the solidification rate, requiring equipment with a high level of technology, making it practically impossible to measure the degree of crystallization. An alternative is using mathematical models applied in computer simulations (CFD), helping to improve and develop new vitrification protocols. This study investigates the vitrification process utilizing the microdroplet method through experimental and numerical analysis.
View Article and Find Full Text PDFRevival of cryopreserved larvae from the important aquaculture species Pacific White Shrimp (Litopenaeus vannamei) using vitrification and ultra-rapid laser warming.
Cryobiology
December 2024
Department of Mechanical Engineering, University of Minnesota, Minneapolis, MN, 55455, USA; Center for Advanced Technologies for the Preservation of Biological Systems (ATP-Bio), University of Minnesota, Minneapolis, MN, 55455, USA.
Cryopreservation of aquatic embryos or larvae is needed to help safeguard genetics from important wild and captive species, increase aquaculture output, and meet the global demand for protein. To this end, the development of a cryopreservation protocol for nauplius larvae of the commercially important aquaculture species Litopenaeus vannamei, or Pacific White Shrimp, was pursued. Toxicity screening was performed using multiple cryoprotective agents (CPA), and a multi-constituent CPA cocktail was developed to achieve reliable vitrification of shrimp larvae encapsulated in 1.
View Article and Find Full Text PDFEvaluating shoot-tip regrowth of 25 Rubus L. species and hybrids after 15 to 20 years of cryopreserved storage.
Cryobiology
November 2024
USDA-ARS, National Clonal Germplasm Repository, 33447 Peoria Rd, Corvallis, OR, 97333, USA, emeritus.
Rubus L. species are pan-global in their distribution and used as food throughout the world. Their fruits, collectively called brambles, come in a variety of colors from black, through various shades of red and yellow to white.
View Article and Find Full Text PDFA microfluidic hanging droplet as a programmable platform for mammalian egg vitrification.
Lab Chip
November 2024
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Egg (oocyte) vitrification is the dominant method for preserving fertility for women of reproductive age. However, the method is typically performed by hand, requiring precise (∼0.1 to 10 μL) and time-sensitive (∼1 s) liquid exchange of cryoprotectants (CPA) around eggs as well as fine handling of eggs (∼100 μm) for immersion into liquid nitrogen (LN).
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!