# Hepatocyte production from ice-free cryopreserved and nanowarmed livers

> **NIH NIH R43** · VITRISTOR LLC · 2021 · $374,262

## Abstract

Hepatocyte-based therapy including cell transplantation, bioartificial and engineered livers are limited by the
inability to produce large quantities of high functioning primary human hepatocytes on demand. In addition, the
pharmaceutical industry could benefit from a technology that can provide pooled donor population and the ability
to control supply with demand for drug metabolism and toxicity testing. The ultimate goal of this project to
develop the technology of vitrification and nanowarming of partial or whole human livers to produce a
broad range of quantities of metabolically active, high quality primary hepatocytes on demand for
therapeutic and pharmaceutical applications.
Vitrification, an ice-free cryopreservation method, shows promise but is currently not applicable to large tissue
and organs due to damaging ice crystal formation during the slow warming. Recently, our group at the University
of Minnesota developed “nanowarming” using iron oxide nanoparticles (IONPs) coupled with radio frequency
(RF) technology to achieve uniform warming rates sufficient to avoid crystallization and cracking in vitrified tissue
and has the ability to scale up to partial and/or whole human organs.
The goal for this Phase I project will be to determine the efficacy of isolating hepatocytes from whole rat
livers that have been vitrified and rewarmed via nanowarming. This project will use the liver’s own native
vascular system to load and unload the vitrification solution (VS) prior to hepatocyte isolation. This allows
homogeneous delivery of the VS to a large number of cells. Our preliminary results show that vitrification of a
whole rat liver and uniform rewarming rates sufficient to avoid crystallization and cracking were achieved.
Loading and unloading of the VS at hypothermic temperatures resulted in high yield, viabiligy and hepatocyte
function. Lastly, nanowarmed vitrified livers showed largely normal architecture, displayed hepatocyte specific
function (indocyanine green uptake) and homogeneous perfusion. This would suggest that large quantities of
viable and functioning cells can be isolated from the nanowarmed vitrified rat liver. The goals of the Phase I
project can be achieved by accomplishing the following Specific Aims:
Specific Aim 1: Determine the efficacy of loading and unloading three different concentrations (7, 8 and
9M) of the vitrification solution (VS) in a rat liver on yield, viability and function of the isolated
hepatocytes.
Specific Aim 2: Determine the IONP concentration that ensures uniform and rapid warming rates
sufficient to avoid crystallization.
Specific Aim 3: Determine the efficacy of vitrifying and nanowarming rat livers for obtaining large
quantities of viable and high functioning hepatocytes.
If successful, the Phase II project will scale-up the technology to porcine livers in collaboration with the Mayo
Clinic. In addition, the project will collaborate with Lonza (world leading hepatocyte supplier) on not-suitable fo...

## Key facts

- **NIH application ID:** 10156435
- **Project number:** 1R43DK126551-01A1
- **Recipient organization:** VITRISTOR LLC
- **Principal Investigator:** Charles Y Lee
- **Activity code:** R43 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $374,262
- **Award type:** 1
- **Project period:** 2021-01-05 → 2023-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10156435

## Citation

> US National Institutes of Health, RePORTER application 10156435, Hepatocyte production from ice-free cryopreserved and nanowarmed livers (1R43DK126551-01A1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10156435. Licensed CC0.

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