Induced Pluripotent Stem Cell Reprogramming via Retroviral Vector Extraction

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Solution Overview

Problem

Current methods for generating human pluripotent stem cells are limited by technical, ethical, and immunological considerations, and there is a need for a reliable source of adult stem cells that can be easily obtained and differentiated for therapeutic and research purposes.

Innovation Solution

The use of retroviral vectors encoding de-differentiation factors such as KLF4, OCT4, SOX2, c-MYC, and NANOG to transform human somatic cells, specifically fibroblasts, into induced pluripotent stem cells (iPS) that are morphologically and genetically similar to embryonic stem cells, allowing for the generation of an enriched population of de-differentiated cells.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If retroviral vectors are used to transform somatic cells into induced pluripotent stem cells, then the generation of patient-specific pluripotent stem cells is achieved, but the risk of chromosomal abnormalities and tumor formation increases

Engineering Contradiction:

Improvegeneration of pluripotent stem cellsVSAvoidchromosomal abnormalities and tumor formation

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent extracts and removes the retroviral vector after successful reprogramming. The method involves selecting for the loss of retroviral expression, thereby eliminating the harmful viral elements while retaining the induced pluripotent stem cell phenotype. This extraction principle resolves the contradiction by removing the source of chromosomal abnormalities and tumor formation risk.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent converts the harmful retroviral vector into a beneficial tool by using it initially for reprogramming, then selecting for its loss. The retroviral vector serves its purpose in transforming somatic cells into induced pluripotent stem cells, and its subsequent removal eliminates the harm. This principle resolves the contradiction by utilizing the vector's reprogramming capability while eliminating its harmful effects.

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

2Reliability

If embryonic stem cells are used for research and therapeutics, then valuable pluripotent cells are obtained, but ethical considerations and immunological rejection risks arise

Engineering Contradiction:

Improvepluripotent stem cell sourceVSAvoidethical considerations and immunological rejection

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent creates induced pluripotent stem cells that are genetically identical to the patient's own somatic cells, effectively copying the patient's genetic material into a pluripotent state. This copying principle resolves the contradiction by providing patient-specific pluripotent cells that avoid ethical issues associated with embryonic stem cells and eliminate immunological rejection risks.

Inventive Principle:

Principle #26Copying

Solution Approach 2:

The patent changes the cellular parameters of somatic cells by introducing de-differentiation factors (KLF4, OCT4, SOX2, c-MYC, NANOG) to transform them into induced pluripotent stem cells. This parameter change resolves the contradiction by converting differentiated patient cells into pluripotent cells with desired properties while maintaining patient-specific genetics.

Inventive Principle:

Principle #35Parameter changes

3Reliability

If de-differentiation factors are introduced into somatic cells, then cell reprogramming to pluripotent state is achieved, but the complexity of the transformation process increases

Engineering Contradiction:

Improvecell reprogramming efficiencyVSAvoidtransformation process complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent segments the reprogramming process into distinct steps: introduction of de-differentiation factors via retroviral vectors, culture and expansion, selection for loss of retroviral expression, and verification of pluripotent state. This segmentation resolves the contradiction by breaking down the complex reprogramming process into manageable, controllable steps.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent incorporates feedback mechanisms through selection for the loss of retroviral expression and verification of pluripotent marker expression. This feedback principle resolves the contradiction by allowing real-time monitoring and selection of successfully reprogrammed cells, thereby improving reprogramming efficiency while maintaining process control.

Inventive Principle:

Principle #23Feedback

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 enables the production of patient-specific pluripotent stem cells that can be used for therapeutic applications, tissue engineering, and research, overcoming limitations of existing technologies by providing an autologous source of stem cells that can differentiate into various cell types without chromosomal abnormalities.

Implementation Method 1

contacting a human somatic cell with at least one retroviral vector comprising polynucleotides encoding at least four de-differentiation factors

Methodology Applied

Scientific EffectRetroviral transduction:

Data Source

PatentUS9005966B2Generation of pluripotent cells from fibroblasts

Publication Date: 2015.04.14 RGT UNIV OF CALIFORNIA

AI summary

Provided are methods and compositions useful for producing and propagating stem cells from fibroblasts.