Charged Microcarrier Tagging for Primary Cell Culture Viability

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Solution Overview

Problem

Current cell culture methods are empirical and inefficient, particularly for growing a wide variety of cells in vitro, as they often rely on trial and error and are inadequate for certain cell types, leading to limited lifespan and loss of cellular characteristics in primary cell lines, and lack accurate models for tissue-specific functions.

Innovation Solution

A method for determining cell culture history using tagged microcarriers and microspheres, allowing for precise measurement and correlation of tag parameters to culture conditions, enabling improved labeling, separation, and analysis of cell units, and the use of microcarriers like Cultispher-G and Cytopore for adherent cell cultures.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Duration of action of stationary object

If conventional cell culture methods are used for primary cells, then cell culture can be established, but the lifespan of primary cell lines is limited and cellular characteristics are lost

Engineering Contradiction:
Improvelifespan of primary cell linesVSAvoidcellular characteristics
Core Design Contradiction:
Duration of action of stationary objectVSReliability

Solution Approach 1:

The invention segments the cell culture system by introducing microcarriers as discrete support structures, allowing primary cells to be cultured in a distributed manner that maintains their characteristics while extending culture lifespan. Each microcarrier acts as an independent unit supporting cell growth.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

Microcarriers serve as an intermediary between the culture medium and primary cells, providing a structured support that maintains cellular characteristics while enabling extended culture duration. The microcarriers mediate the interaction between cells and culture conditions.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Adaptability or versatility

If empirical trial and error methods are used for cell culture, then various cell types can be attempted, but the process is inefficient and inadequate for certain cell types

Engineering Contradiction:
Improveability to grow wide variety of cellsVSAvoidefficiency of cell culture process
Core Design Contradiction:
Adaptability or versatilityVSProductivity

Solution Approach 1:

The microcarrier system provides a universal platform that can support the culture of diverse cell types including primary cells, cell lines, and stem cells. The same microcarrier technology serves multiple cell culture applications, eliminating the need for type-specific culture methods.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Solution Approach 2:

The invention changes key culture parameters by introducing microcarriers with specific physical and chemical properties (surface charge, porosity, material composition) that optimize cell attachment and growth, transforming empirical trial-and-error into a systematic approach with controllable parameters.

Inventive Principle:
Principle #35Parameter changes

3Duration of action of stationary object

If continuous cell lines are established to extend culture lifespan, then cells can be cultured indefinitely, but tissue-specific characteristics are lost

Engineering Contradiction:
Improveculture lifespanVSAvoidtissue-specific characteristics
Core Design Contradiction:
Duration of action of stationary objectVSReliability

Solution Approach 1:

The microcarriers are pre-prepared with optimal surface properties and charged tags before cell culture initiation, creating a stable environment that maintains tissue-specific characteristics from the outset rather than allowing drift over time. This preliminary preparation prevents the loss of cellular characteristics during extended culture.

Inventive Principle:
Principle #10Preliminary action

4Strength

If tags are coated with proteins to improve binding, then attachment to microcarriers is enhanced, but measurement and analysis become more complex

Engineering Contradiction:
Improvebinding strength of tagsVSAvoidcomplexity of tag analysis
Core Design Contradiction:
StrengthVSDevice complexity

Solution Approach 1:

The invention extracts and removes proteins from the tag coating, using instead a simplified charged microsphere design that relies on electrostatic interactions. This extraction eliminates the complexity of protein-based tagging systems while maintaining effective binding to microcarriers.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention changes the tagging mechanism from protein-based to charge-based, using microspheres with defined surface charges that simplify the physical and chemical parameters of the system. This parameter change reduces analytical complexity while maintaining binding effectiveness.

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 allows for the prolonged viability and accurate representation of primary cells, improved reliability of cell-based experiments, and systematic sampling of multiple culture conditions, enhancing the precision and predictive power of cellular processes and differentiation.

Implementation Method 1

a microcarrier having a positive charge conjugated to a negatively charged tag

Methodology Applied
Scientific EffectElectrostatic attraction: Electrostatics

Data Source

PatentEP1917349B1Complex comprising a microcarrier having a positive charge conjugated to a negatively charged tag which tag is microsphere, which microsphere is not coated with proteins and uses thereof.
Publication Date: 2013.09.25 PLASTICELL LTD
  • EP1917349B1 patent drawingFigure 1
  • EP1917349B1 patent drawingFigure 2
  • EP1917349B1 patent drawingFigure 3

AI summary

The present invention relates in one aspect to a method for determining the cell culture history of a cell unit labelled with more than one type of tag comprising the steps of: (a) measuring one or more parameters of each tag that is used to label the cell unit; (b) identifying each tag in the cell unit; and (c) correlating the identity of each tag to the identity of the cell unit and/or the specific cell culture conditions to which the cell unit has been exposed.