Lymphocyte Cryopreservation With Bicarbonate Pre-Washing

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Solution Overview

Problem

Existing washing and cryopreservation solutions for lymphocytes, such as T cells and NK cells, do not adequately maintain high cell recovery and survival rates after freezing and thawing, leading to time loss and reduced efficacy in cell therapies.

Innovation Solution

A method involving washing lymphocytes with a bicarbonate ion-containing physiological aqueous solution, followed by cryopreservation in a solution containing dextran, glucose, DMSO, and human serum albumin, significantly improves cell recovery and survival rates after thawing.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional cryopreservation solutions are used, then cells can be stored and transported, but cell recovery rate and survival rate decrease after thawing

Engineering Contradiction:

Improvecell recovery rateVSAvoidharmful components in cryopreservation solution

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention extracts and removes harmful components from the cryopreservation solution by implementing a washing step before freezing. This eliminates toxic medium components while preserving the essential cryopreservation function, thereby improving cell recovery rate and survival rate after thawing without compromising storage capability.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention performs a preliminary washing action before the cryopreservation process to remove harmful components from the culture medium. This preliminary cleaning step prevents harmful substances from being present during freezing and storage, ensuring higher cell viability upon thawing while maintaining the ability to store and transport cells.

Inventive Principle:

Principle #10Preliminary action

2Object-affected harmful factors

If washing is performed to remove harmful components, then cell purity improves, but time loss and reduced cell recovery rate occur

Engineering Contradiction:

Improveharmful componentsVSAvoidwashing time

Core Design Contradiction:

Object-affected harmful factorsVSLoss of time

Solution Approach 1:

The invention optimizes the washing parameters by using a simplified bicarbonate buffer solution instead of complex conventional washing protocols. This parameter change reduces washing time while effectively removing harmful components, thereby improving cell purity without significant time loss or reduction in cell recovery rate.

Inventive Principle:

Principle #35Parameter changes

3Reliability

If multiple medium components are used in cryopreservation solution, then cell protection during freezing is improved, but harmful substances increase

Engineering Contradiction:

Improvecell protection during freezingVSAvoidtoxic medium components

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The invention extracts harmful medium components from the cryopreservation formulation by implementing a pre-freezing washing step. This allows the use of essential protective components during freezing while removing toxic substances, thereby maintaining cell protection during freezing without exposing cells to harmful medium components.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention segments the cryopreservation process into two distinct phases: a washing phase to remove harmful components, and a freezing phase using a simplified solution. This segmentation allows optimal protection during freezing while eliminating toxic substances, resolving the contradiction between cell protection and harmful component reduction.

Inventive Principle:

Principle #1Segmentation

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

The proposed method ensures high cell recovery and survival rates, even after prolonged storage at room temperature, making lymphocytes more suitable for transplantation.

Implementation Method 1

a bicarbonate ion-containing physiological aqueous solution

Methodology Applied

Scientific EffectOsmotic pressure: Osmotic Pressure

Implementation Method 2

bicarbonate ion-containing physiological aqueous solution

Methodology Applied

Scientific EffectBuffering capacity:

Implementation Method 3

cryopreservation solution containing a physiological aqueous solution, preferably a bicarbonate ion-containing physiological aqueous solution

Methodology Applied

Scientific EffectCryoprotection:

Implementation Method 4

cryopreservation

Methodology Applied

Scientific EffectFreezing: Freezing

Data Source

PatentEP4621046A1Production and cryopreservation method for lymphocytes, cleansing liquid and cryopreservation liquid for same, and preparation of lymphocytes for transplantation from cryopreserved lymphocytes

Publication Date: 2025.09.24 HEALIOS KK

AI summary

The present invention provides a method for washing lymphocytes, including washing lymphocytes with a bicarbonate ion-containing physiological aqueous solution; a method for cryopreserving lymphocytes, including suspending and freezing lymphocytes obtained by the method in a cryopreservation solution containing a physiological aqueous solution; a washing solution for lymphocytes, including a bicarbonate ion-containing physiological aqueous solution; a cryopreservation solution for lymphocytes, containing a bicarbonate ion-containing physiological aqueous solution, dextran, glucose, DMSO, and human serum albumin; a lymphocyte-containing composition, containing lymphocytes treated with a bicarbonate ion-containing physiological aqueous solution and the aforementioned cryopreservation solution; a kit for preparing lymphocytes for transplantation, containing the lymphocyte-containing composition and a diluent for transplantation containing a bicarbonate ion-containing physiological aqueous solution, and the like.