A CryoDAO Project

High-Viscosity Solutions to Mitigate Perfusion Impairment

High-Viscosity Solutions to Mitigate Perfusion Impairment
Aschwin De Wolf
Funding Goals



Project Overview

Human cryopreservation is exclusively practiced as a post-mortem procedure. This leads to perfusion under ischemic conditions in a relevant number of cases. Ischemic conditions lead to perfusion impairment and in turn low quality preservation. We propose a novel high-viscosity cryoprotective agent (CPA) solution to mitigate ischemia-induced perfusion impairment and relevantly improve the quality of the average cryopreservation case.

We propose to research, develop, and validate a High-Viscosity and/or High-Tonicity CPA Solutions. One protocol that was shown some preliminary benefits was to modify the cryoprotectant carrier solution to have high viscosity. In this research we will compare vitrification solutions:

- normal viscosity

- moderately increased viscosity

- high viscosity 

Viscosity modification will be done with trehalose and Polyvinylpyrrolidone (PVP). 

The project plans to establish whether high-viscosity cryoprotectants can (partly) reverse ischemia-induced perfusion impairment and ice formation.


We aim to validate this solution in an in-situ rat brain cryopreservation model with elimination of ice formation as a clinical endpoint. Our model provides for a non-ischemic control, an experimental group of 2 hours of normothermic ischemia, and a 2 normothermic ischemia experimental group in which we administer the solution.

Expected Results:

We expect to see significant improvement in mitigation of perfusion impairment and reduction of ice formation after cryopreservation in the group that was perfused with the novel CPA solution. Successful development and validation of such a solution would strongly streamline and simplify cryonics perfusion procedures and even hold promise for complex tissue and organ preservation.


Lead: Aschwin de Wolf (AdW), Advanced Neural Biosciences, NY
Staff Scientists at Advanced Neural Biosciences

Next Steps:

AdW lab has done extensive prior work in applied and translational cryopreservation research and in-situ rat brain cryopreservation model, as well as evaluation of cryopreservation quality.

- Milestone 1: Creation of High-viscosity solutions and initial validation. If results are promising, continue with milestone 2.

- Milestone 2: Detailed validation and quality assessment of preservation quality under ischemic conditions.


High-Viscosity Solutions to Mitigate Perfusion Impairment

- Creation of one or more High-viscosity solution(s) and initial validation - USD 25,000

- Detailed validation and quality assessment of preservation quality under ischemic conditions - USD 25,000

Total: 6-month personnel + running costs - USD 50,000

No overhead


  • High impact for human cryopreservation
  • Comparatively low risk  
  • Solid evidence supporting their approach
  • All required technology established in lab
  • Follow on project has commercial potential, founding of company is possible and supported by PI  
  • Patent would have commercial potential
  • Pre-clinical applications (e.g. organ preservation for research), as well as applications for organ and human cryopreservation.
  • Comparatively low funding required
  • No university overhead, private lab without TTO/third party approval required 


  • Short term commercialization might not be easy, but relevant long-term potential
  • Technical risk

Outcome of the evaluation and recommendation:

A total of X evaluators independently scored the project proposal on different categories as either: (1) Outstanding, (2) Strong, (3) Satisfactory, (4) Weak, (5) Unacceptable, (N/A) Not enough information provided, or (N/A) Not my area of expertise. This is a summary of the results:

  • Novelty and Impact: (2) Strong (1/X evaluators)
  • Feasibility and Data: (1) Outstanding (1/X evaluators)
  • Relevance to cryopreservation: (1) Outstanding (1/X evaluators)
  • Science Team: (2) Strong (1/X evaluators)
  • Market Advantage: (2) Strong (1/X evaluators)
  • IP-NFT Potential: (2) Strong (1/X evaluators)

All the evaluators consider the project worth funding by the CryoDAO community.

Mechanism of Funding:

This proposal recommends CryoDAO commits funding via an IP-NFT. No TTO or similar third party approval is needed.


While Human Cryopreservation holds tremendous potential it is still a majorly underfunded sector of the longevity and life extension ecosystem. Even comparatively modest funding amounts can lead to significant and relevant impact. Ischemia-induced perfusion impairment is one of the major issues as a relevant number of cryopreservation cases are done under different degrees of ischemia, primarily due to logistical complexities. Similar problems arise in organ preservation cases as well.


Positive results could be the base for novel IP regarding formulation of solutions and delivery methods to improve cryopreservation quality, and potential complex tissue and organ preservation at higher temperature, significantly. Preliminary data shows that a few of the tested solutions have beneficial effects on preservation quality. Results from the proposed study will provide critical preclinical IP to support commercializing a High-Viscosity Solution to Mitigate Perfusion Impairment under ischemic conditions, which the investigator(s) intend to do with support from CryoDAO.

Funding can be provided in two milestones to reduce the overall risk.