Non-invasive Genetic Integrity Assessment of Pluripotent Stem Cells

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Solution Overview

Problem

Current methods for assessing the genetic integrity of pluripotent stem cells in culture are invasive, time-consuming, and lack consensus on distinguishing between potentially carcinogenic mutations and benign polymorphisms, posing a risk for malignant transformation and therapeutic efficacy.

Innovation Solution

A non-invasive method using hyper-recurrent sequences as biomarkers to detect genetic abnormalities in cell culture supernatants through techniques like droplet digital-PCR, allowing for rapid assessment and isolation of stem cells free from genetic defects.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If invasive methods are used to assess genetic integrity of pluripotent stem cells, then measurement precision is improved, but productivity deteriorates and loss of time increases

Engineering Contradiction:
Improvegenetic integrity assessment accuracyVSAvoidcell assessment throughput
Core Design Contradiction:
Measurement precisionVSProductivity

Solution Approach 1:

The invention extracts and analyzes cell-free nucleic acids from culture supernatants rather than requiring direct cell manipulation. This extraction approach allows genetic integrity assessment without invasive cell processing, maintaining productivity while achieving measurement precision through analysis of circulating nucleic acid markers that reflect cellular genetic status.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention uses cell-free nucleic acids in culture supernatants as an intermediary medium to assess genetic integrity. Instead of directly analyzing cells through invasive methods, the nucleic acid markers in the supernatant serve as mediators that convey genetic information, enabling non-invasive assessment that maintains both precision and productivity.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Measurement precision

If conventional genetic analysis methods are used, then measurement precision is improved, but device complexity and difficulty of detecting and measuring increase

Engineering Contradiction:
Improvegenetic abnormality detection accuracyVSAvoidanalysis system complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The invention simplifies the analysis system by extracting and analyzing only cell-free nucleic acids from supernatants rather than requiring complex whole-cell genomic analysis. This extraction strategy reduces device complexity while maintaining measurement precision by focusing on specific genetic markers that indicate chromosomal abnormalities.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention applies local quality analysis by examining specific genetic regions and markers within the nucleic acids rather than analyzing the entire genome. This targeted approach to detecting genetic abnormalities in supernatants reduces the complexity of the detection system while maintaining high measurement precision for clinically relevant genetic changes.

Inventive Principle:
Principle #3Local quality

3Reliability

If comprehensive genetic screening is performed, then reliability is improved, but loss of time and productivity deteriorate

Engineering Contradiction:
Improvetherapeutic safety assuranceVSAvoidscreening duration
Core Design Contradiction:
ReliabilityVSLoss of time

Solution Approach 1:

The invention extracts and analyzes only the most clinically relevant genetic markers from cell-free nucleic acids in supernatants, rather than performing comprehensive whole-genome screening. This targeted extraction approach maintains reliability for detecting critical chromosomal abnormalities while significantly reducing screening time and improving productivity.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention applies partial action by performing focused genetic screening on specific high-risk genetic markers rather than comprehensive analysis of all genetic material. This partial screening approach provides sufficient reliability for therapeutic safety by detecting the most critical abnormalities, while reducing time loss and improving overall productivity.

Inventive Principle:
Principle #16Partial or excessive action

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

Enables quick, inexpensive, and reliable detection of genetic abnormalities, ensuring the quality and safety of pluripotent stem cells for clinical use by identifying hyper-recurrent sequences in supernatants, thereby reducing the risk of malignant transformation and improving therapeutic outcomes.

Implementation Method 1

A non-invasive method using hyper-recurrent sequences as biomarkers to detect genetic abnormalities in cell culture supernatants through techniques like droplet digital-PCR

Methodology Applied
Scientific EffectPolymerase chain reaction (PCR):

Data Source

PatentEP3347487B1Non-invasive methods for assessing genetic integrity of pluripotent stem cells
Publication Date: 2021.07.14 INST NAT DE LA SANTE & DE LA RECHERCHE MEDICALE (INSERM)
  • EP3347487B1 patent drawingFigure 1~2
  • EP3347487B1 patent drawingFigure 3
  • EP3347487B1 patent drawingFigure 4

AI summary

The present invention relates to a novel non-invasive method for assessing pluripotent stem cells quality in culture. More specifically, the present invention relates to a non-invasive method for assessing genetic integrity (such as the presence of CNVs) of pluripotent stem cells in culture, by assessing cell-free nucleic acids in the supernatant of the cell culture.