Monocyte Proliferation via Cytokine Culture Medium

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Solution Overview

Problem

Current methods for producing dendritic cell vaccines are inefficient and burdensome, requiring multiple apheresis sessions and struggling to achieve sufficient cell numbers due to low monocyte proliferation rates, which can lead to prolonged treatment times and patient discomfort.

Innovation Solution

A method involving the use of a cytokine-rich culture medium containing GM-CSF, IFN-γ, and Flt-3L to efficiently proliferate monocytes in vitro, allowing for the production of sufficient dendritic cells without the need for repeated cell cultures and reducing patient burden.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If conventional apheresis method is used to collect monocytes, then monocytes can be obtained, but the apparatus is expensive and requires advanced skills to operate

Engineering Contradiction:
Improvemonocyte collectionVSAvoidoperation complexity
Core Design Contradiction:
Quantity of substanceVSEase of operation

Solution Approach 1:

The patent replaces expensive, complex apheresis apparatus with simple, disposable materials including centrifuge tubes, culture plates, and standard laboratory equipment. The method uses conventional centrifugation and culture techniques that require minimal specialized equipment, making the process accessible to ordinary laboratories without expensive apheresis machines.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent divides the monocyte collection process into separate, manageable steps: blood collection by simple venipuncture, centrifugation to separate mononuclear cells, and culture to enrich monocytes. This segmentation allows each step to be performed with simple, standard equipment rather than requiring a single complex apheresis apparatus.

Inventive Principle:
Principle #1Segmentation

2Quantity of substance

If conventional apheresis method is used to collect monocytes, then monocytes can be obtained, but multiple apheresis sessions are required to achieve sufficient cell numbers

Engineering Contradiction:
Improvemonocyte collectionVSAvoidtreatment time
Core Design Contradiction:
Quantity of substanceVSLoss of time

Solution Approach 1:

The patent performs preliminary monocyte enrichment through in vitro culture before vaccine production begins. By culturing monocytes for 3-7 days with appropriate cytokines (GM-CSF, M-CSF, or GM-CSF plus IL-4), the method pre-concentrates monocytes from the initial blood sample, eliminating the need for repeated apheresis sessions to accumulate sufficient cell numbers.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent establishes continuous monocyte proliferation through culture conditions that maintain high monocyte viability and promote division. The use of cytokine-containing culture media creates a continuous growth environment, allowing monocytes to multiply over several days rather than requiring discrete, repeated collection sessions.

Inventive Principle:
Principle #20Continuity of useful action

3Quantity of substance

If conventional culture method is used to differentiate monocytes into dendritic cells, then differentiation can occur, but sufficient cell numbers cannot be obtained

Engineering Contradiction:
Improvedendritic cell productionVSAvoidcell proliferation rate
Core Design Contradiction:
Quantity of substanceVSProductivity

Solution Approach 1:

The patent optimizes culture parameters including cytokine concentrations (GM-CSF at 100-500 ng/mL, M-CSF at 100-500 ng/mL, or GM-CSF plus IL-4), culture duration (3-7 days for monocyte enrichment followed by differentiation), and cell density to maximize both monocyte proliferation and subsequent dendritic cell differentiation efficiency, achieving sufficient cell numbers for vaccine production.

Inventive Principle:
Principle #35Parameter changes

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

This approach enables the rapid and efficient production of a sufficient number of dendritic cells for vaccine production, reducing treatment time and minimizing patient discomfort while maintaining high cell viability and antigen-presenting ability.

Implementation Method 1

a method involving the use of a cytokine-rich culture medium containing GM-CSF, IFN-γ, and Flt-3L to efficiently proliferate monocytes in vitro

Methodology Applied
Scientific EffectCytokine-mediated cell proliferation:

Data Source

PatentEP2749639B1Proliferating agent for monocyte, culture medium for proliferating monocyte, method for producing monocyte, method for producing dendritic cell, and method for producing dendritic cell vaccine
Publication Date: 2016.10.12 HAKUSHINKOUSEIKAI FOUND
  • EP2749639B1 patent drawingFigure 1A~1B
  • EP2749639B1 patent drawingFigure 2
  • EP2749639B1 patent drawingFigure 3A~3C

AI summary

The purpose of the present invention is to provide a means for proliferating a monocyte with high efficiency and in a simple manner. The present invention provides a proliferating agent for a monocyte, which consists of at least one component selected from Flt-3L, IL-3 and IFN-γ and can be used before a treatment for differentiation of a monocyte into a dendritic cell. The present invention also provides a culture medium for use in the proliferation of a monocyte, which contains at least one component selected from Flt-3L, IL-3 and IFN-γ and can be used before a treatment for differentiation of a monocyte into a dendritic cell. The culture medium for use in the proliferation of a monocyte according to the present invention may contain GM-CSF.