Enucleated Cell Cryopreservation for Pure Therapeutic Delivery

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Existing large-scale manufacturing techniques for enucleated cells result in the presence of nucleated parent cells, undermining the benefits of enucleated cell platforms, such as unwanted gene transfer and limited lifespan in vivo.

Innovation Solution

Cryopreservation of enucleated cells in compositions like liquid nitrogen or CryoStor® media, followed by thawing and reconstitution, enhances the delivery efficiency of therapeutic agents, maintaining or increasing the amount of agent delivery compared to non-cryopreserved cells.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If large-scale manufacturing techniques are used to produce enucleated cells, then productivity is improved, but manufacturing precision deteriorates due to presence of nucleated parent cells

Engineering Contradiction:

Improvelarge-scale manufacturing capabilityVSAvoidpurity of enucleated cells

Core Design Contradiction:

ProductivityVSManufacturing precision

Solution Approach 1:

The patent applies preliminary action by performing enucleation on nucleated cells before cryopreservation. This ensures that only pure enucleated cells are stored and subsequently used for therapeutic agent delivery, eliminating the risk of nucleated parent cell contamination in the final product while maintaining large-scale manufacturing capability

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent extracts the nucleus from parent cells to create enucleated cells. This extraction process removes the genetic material that could cause unwanted gene transfer, while preserving the cytoplasmic components necessary for therapeutic function. The enucleated cells are then purified to remove any remaining nucleated cells before cryopreservation

Inventive Principle:

Principle #2Taking out (Extraction)

2Reliability

If enucleated cells are used as therapeutic platform, then reliability is improved by avoiding gene transfer, but manufacturing precision worsens due to difficulty in complete enucleation

Engineering Contradiction:

Improvesafety against gene transferVSAvoidcompleteness of enucleation

Core Design Contradiction:

ReliabilityVSManufacturing precision

Solution Approach 1:

The patent replaces mechanical/enzymatic enucleation methods with a chemical approach using a cell-permeable compound that induces nuclear envelope breakdown. This chemical method more effectively removes nuclear material while preserving cytoplasmic integrity, achieving more complete enucleation and reducing residual nuclear DNA that could cause gene transfer

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent changes the chemical parameters of the enucleation process by using specific cell-permeable compounds at controlled concentrations and exposure times. This allows optimization of the enucleation reaction to achieve complete nuclear removal while maintaining cell viability and therapeutic function, thereby improving both reliability and manufacturing precision

Inventive Principle:

Principle #35Parameter changes

3Duration of action of stationary object

If enucleated cells are cryopreserved for long-term storage, then duration of action is improved, but manufacturing precision may worsen due to potential cell damage

Engineering Contradiction:

Improvestorage durationVSAvoidcell integrity after thawing

Core Design Contradiction:

Duration of action of stationary objectVSManufacturing precision

Solution Approach 1:

The patent utilizes controlled phase transitions during cryopreservation and thawing to protect cell integrity. By carefully controlling the freezing and thawing rates, the patent prevents ice crystal formation that could damage enucleated cells, thereby maintaining cell integrity after long-term storage and subsequent thawing

Inventive Principle:

Principle #36Phase transitions

Solution Approach 2:

The patent applies beforehand cushioning by using cryoprotectant agents before freezing the enucleated cells. These protective agents prevent cellular damage during the freezing process by reducing ice crystal formation and stabilizing cell membranes, ensuring that cells maintain their integrity and therapeutic function after long-term cryopreservation and subsequent thawing

Inventive Principle:

Principle #11Beforehand cushioning (Prior cushioning)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

Cryopreserved enucleated cells effectively deliver therapeutic agents, such as viruses and proteins, at higher titers or secretion levels, suitable for treating conditions like cancer, by maintaining or enhancing the therapeutic efficacy.

Implementation Method 1

CryoStor® media has been shown to stabilize enucleated cells during cryopreservation and after thawing

Methodology Applied

Scientific EffectCryopreservation: Freezing

Implementation Method 2

Cryopreserved enucleated cells effectively deliver therapeutic agents, such as viruses and proteins, at higher titers or secretion levels

Methodology Applied

Scientific EffectCellular delivery mechanism:

Data Source

PatentUS20260021053A1Manufacturing systems and methods for cellular therapeutic platforms

Publication Date: 2026.01.22 RGT UNIV OF CALIFORNIA

AI summary

Described herein are methods for obtaining enucleated cells from nucleated cells. Also described herein are methods for cell processing, including providing a composition containing nucleated cells and enucleating at least a portion of the nucleated cells to produce an enucleated cell fraction. Also described herein are methods for cell processing, including expressing the heterologous gene product. Also provided are pharmaceuticals compositions comprising an enucleated cell.