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Bulk Droplet Vitrification for Primary Hepatocyte Preservation
Published on: October 25, 2019
Microencapsule technique protects hepatocytes from cryoinjury.
Tomokazu Kusano1, Takeshi Aoki, Daisuke Yasuda
1General and Gastroenterological Surgery, Showa University School of Medicine, Tokyo, Japan.
Summary
Microencapsulation protects hepatocytes from cryoinjury, preserving cell viability and hepatocyte nuclear factor (HNF) levels. This technique is vital for establishing effective hepatocyte banks for transplantation.
Area of Science:
- Regenerative Medicine
- Cell Biology
- Biotechnology
Background:
- Hepatocyte transplantation offers an alternative to whole liver transplants.
- Establishing a functional hepatocyte bank is crucial for this procedure.
- Microencapsulation is a promising method for hepatocyte cryopreservation.
Purpose of the Study:
- To investigate the prevention of cryoinjury in microencapsulated hepatocytes.
- To assess the efficacy of alginate-poly L-lysine microencapsulation for hepatocyte cryopreservation.
- To evaluate the preservation of hepatocyte function post-cryopreservation.
Main Methods:
- Hepatocytes were isolated using EDTA/collagenase digestion.
- Cells were microencapsulated in alginate-poly L-lysine and cryopreserved in liquid nitrogen.
- Cryomicroscopy, ultrastructural examination, and RT-PCR for hepatocyte nuclear factor (HNF) were employed.
Main Results:
- Alginate microencapsulation prevented physical damage from extracellular ice crystals.
- Intracellular hepatocyte structure was maintained post-cryopreservation.
- Hepatocyte nuclear factor (HNF) levels remained stable, indicating preserved function.
Conclusions:
- The microencapsulation technique effectively protects hepatocytes from cryoinjury.
- This method ensures cell viability and function after cryopreservation.
- The technique is suitable for developing hepatocyte banks for clinical applications.
