BC2LCN Lectin for Undifferentiated Stem Cell Detection

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Solution Overview

Problem

Current methods for determining the differentiation status of stem cells lack specificity and reliability, particularly in distinguishing between undifferentiated and differentiated states, which is crucial for ensuring the quality and safety of stem cell therapies.

Innovation Solution

The use of BC2LCN lectin, which specifically recognizes the sugar chain structures Fucα1-2Galβ1-3GlcNAc and Fucα1-2Galβ1-3GalNAc, allows for accurate differentiation status evaluation and isolation of undifferentiated stem cells through fluorescent labeling and detection, enabling effective quality control and contamination assessment.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional methods are used to determine differentiation status of stem cells, then the process is simple, but the measurement precision and reliability are insufficient

Engineering Contradiction:
Improvedifferentiation status detection accuracyVSAvoiddetection system complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent introduces lectin as an intermediary substance that specifically binds to sugar chain structures on the cell surface. This lectin acts as a mediator between the detection system and the stem cells, enabling specific recognition of undifferentiated cells through their unique sugar chain markers without requiring complex intracellular analysis

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent employs fluorescent labeling of lectin to enable visual detection and sorting of undifferentiated stem cells. The fluorescent marker provides a clear optical signal that changes based on lectin binding to specific sugar chains, allowing for precise identification and separation of cell populations based on their differentiation status

Inventive Principle:
Principle #32Color changes

2Reliability

If stem cells are not properly evaluated for undifferentiated cells, then the process is quick, but the reliability of therapy and safety are compromised

Engineering Contradiction:
Improvestem cell therapy reliabilityVSAvoidquality evaluation time
Core Design Contradiction:
ReliabilityVSLoss of time

Solution Approach 1:

The patent performs quality evaluation of stem cells before transplantation by detecting undifferentiated cells using lectin binding. This preliminary assessment ensures that only appropriately differentiated cells are used for therapy, preventing tumorgenesis while maintaining efficient workflow through a streamlined detection protocol

Inventive Principle:
Principle #10Preliminary action

3Measurement precision

If general lectin is used for cell detection, then the method is simple, but the specificity for undifferentiated stem cells is low

Engineering Contradiction:
Improveundifferentiated cell detection specificityVSAvoiddetection method simplicity
Core Design Contradiction:
Measurement precisionVSEase of manufacture

Solution Approach 1:

The patent exploits the local quality difference in sugar chain structures between undifferentiated and differentiated cells. Undifferentiated stem cells possess specific sugar chain markers (such as sialyl Lewis x) that are absent or reduced in differentiated cells. The lectin is designed to specifically recognize these localized molecular features on the cell surface, providing high detection specificity

Inventive Principle:
Principle #3Local quality

Solution Approach 2:

The patent changes the detection parameter from general cell surface markers to specific sugar chain structures. By targeting the biochemical composition (sugar chains) rather than protein markers, the method achieves higher specificity for undifferentiated stem cells while maintaining procedural simplicity through direct lectin binding assays

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

BC2LCN lectin provides high specificity and affinity for undifferentiated stem cells, enabling reliable detection and separation, thereby improving the quality control of stem cells and reducing the risk of tumorgenesis in therapies.

Implementation Method 1

BC2LCN lectin, which specifically recognizes the sugar chain structures Fucα1-2Galβ1-3GlcNAc and Fucα1-2Galβ1-3GalNAc

Methodology Applied
Scientific EffectLectin-sugar chain recognition: Adsorption

Implementation Method 2

fluorescent labeling and detection

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP2821498B1Method for isolating undifferentiated stem cells
Publication Date: 2018.12.12 FUJIFILM WAKO PURE CHEMICAL CORP
  • EP2821498B1 patent drawingFigure 1A~1B
  • EP2821498B1 patent drawingFigure 2A~2D
  • EP2821498B1 patent drawingFigure 3A~3D

AI summary

Provided are a method for accurately evaluating the differentiation status of stem cells by selectively staining only stem cells in an undifferentiated state, and a method for positively isolating only stem cells in an undifferentiated state. Specifically provided is a method for determining differentiation of a cell comprising a step of contacting a test cell with a probe comprising protein (A) or (B) below and a step of detecting the presence of binding of the probe to the test cell. The method for determining differentiation of a cell is capable of detecting the presence or absence of an undifferentiated stem cell in test cells by using a probe that specifically reacts with undifferentiated stem cells and detecting the presence of bonding to the test cell. (A) A protein comprising an amino acid sequence shown in SEQ ID NO: 1 and recognizing a sugar chain structure of "Fuc±1-2Gal²1-3GlcNAc" and/or "Fuc±1-2Gal²1-3GalNAc;" and (B) a protein comprising an amino acid sequence showing 80% or more similarity to the amino acid sequence shown in SEQ ID NO: 1 and recognizing a sugar chain structure of "Fuc±1-2Gal²1-3GlcNAc" and/or "Fuc±1-2Gal²1-3GalNAc."