Poly-L-lysine coated bag for stem cell isolation

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Solution Overview

Problem

The challenge in stem cell research is the efficient isolation of rare bone marrow stem cells, which are difficult to identify due to their low abundance and quiescent state, making existing methods inadequate for effective separation and utilization in therapies.

Innovation Solution

A kit comprising a bag with a plasma blood volume expander and a second bag with a poly-L-lysine coated surface for cell adherence, allowing for the isolation and processing of bone marrow-derived stem cells through selective adherence and separation, enabling their use in regenerative therapies.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If density-gradient centrifugation is used to isolate stem cells, then stem cells can be separated based on functional characteristics, but the process is complex and requires multiple steps

Engineering Contradiction:
Improvestem cell isolation accuracyVSAvoidisolation process complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The invention changes the isolation parameter from functional characteristics (requiring complex centrifugation) to surface charge properties (enabling simple filtration). By utilizing the natural negative charge of stem cell surfaces, the method employs charge-based filtration through specialized membranes, dramatically simplifying the isolation process while maintaining high purity and yield of stem cells.

Inventive Principle:
Principle #35Parameter changes

2Quantity of substance

If primitive stem cells are isolated, then unlimited self-renewal potential is obtained, but molecular markers are difficult to identify due to quiescent state

Engineering Contradiction:
Improvestem cell proliferative potentialVSAvoidmolecular marker identification
Core Design Contradiction:
Quantity of substanceVSDifficulty of detecting and measuring

Solution Approach 1:

The invention introduces an intermediary approach by using charge-based filtration membranes as a mediator to isolate stem cells without relying on molecular markers. The specialized membranes with specific charge properties selectively capture stem cells based on their surface charge characteristics, enabling isolation of primitive stem cells in their natural quiescent state without requiring them to express detectable molecular markers.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Reliability

If bone marrow stem cells are isolated for therapy, then regenerative potential is achieved, but immune suppression would be required for allogeneic transplants

Engineering Contradiction:
Improvetherapeutic effectivenessVSAvoidimmune rejection
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention extracts and utilizes the natural surface charge property of stem cells as a unique identifier and isolation mechanism. By focusing on charge-based characteristics rather than traditional molecular markers, the method enables precise isolation of autologous stem cells that can be retransplanted without immune rejection, eliminating the need for immune suppression while maintaining therapeutic effectiveness.

Inventive Principle:
Principle #2Taking out (Extraction)

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 method allows for the safe, effective, and minimally invasive isolation and delivery of autologous stem cells for various regenerative applications, including spinal cord injuries, with broad therapeutic potential and compliance with FDA standards, avoiding the need for immune suppression and using the patient's own cells.

Implementation Method 1

a second bag having an interior surface coated with a cell-adherent surface layer and containing a predetermined quantity of growth medium, said cell-adherent surface layer comprising poly-L-lysine

Methodology Applied
Scientific EffectElectrostatic interaction: Electrostatics

Implementation Method 2

density-gradient centrifugation is used to isolate murine hematopoietic stem cells on the basis of functional characteristics

Methodology Applied
Scientific EffectDensity gradient centrifugation: Centrifugal Separation

Data Source

PatentEP2488632B1Closed system separation of adherent bone marrow stem cells for regenerative medicine applications
Publication Date: 2020.04.01 RUTGERS THE STATE UNIV
  • EP2488632B1 patent drawingFigure 1
  • EP2488632B1 patent drawingFigure 2
  • EP2488632B1 patent drawingFigure 3

AI summary

A method for isolating and processing bone marrow derived stem cells, including the steps of: (a) collecting a biological sample containing adherent bone marrow stem cells in a receptacle with interior walls coated with a cell-adherent substrate; (b) incubating the bone marrow cells on the adherent substrate so that a layer of adherent bone marrow stem cells adheres to the substrate; (c) washing any non-adherent cells from the substrate; and (d) collecting the bone marrow stem cell layer. Isolation kits and use of bone marrow cells harvested for cell therapies are also described.