Adipose Stem Cell Culture Using CORM-2 and N-acetyl-L-cysteine

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Solution Overview

Problem

Human adipose tissue-derived multipotent stem cells with high proliferation rates and ability to maintain an undifferentiated state for a long period are difficult to isolate and differentiate into various cell types, limiting their application in treating diseases such as osteoarthritis, osteoporosis, and diabetes.

Innovation Solution

A method involving culturing human adipose tissue-derived pellets in a medium containing N-acetyl-L-cysteine and CORM-2 to maintain stem cells in an undifferentiated state, with specific differentiation protocols using DMEM, α-MEM, and other media compositions to induce differentiation into nerve cells, cartilage cells, osteogenic cells, fat cells, and insulin-releasing pancreatic beta-cells, and forming cellular therapeutic agents.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If adult stem cells are isolated from various human organs, then they can differentiate into specific tissues, but they are very rarely present and difficult to culture without inducing differentiation

Engineering Contradiction:

Improvedifferentiation capabilityVSAvoidculture difficulty

Core Design Contradiction:

Adaptability or versatilityVSEase of manufacture

Solution Approach 1:

The patent changes the cultural parameters by using specific media compositions (containing N-acetyl-L-cysteine, ascorbic acid, calcium, rEGF, BPE, insulin, and hydrocortisone) and controlling culture conditions (37°C, 5% CO2) to maintain stem cells in an undifferentiated state, enabling long-term culture without spontaneous differentiation

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent uses sphere formation as an intermediary mechanism to maintain stem cells in an undifferentiated state. By culturing cells in suspension to form spheres, the cells are protected from differentiation and can be maintained indefinitely, serving as a bridge between isolation and therapeutic application

Inventive Principle:

Principle #24Intermediary (Mediator)

2Duration of action of stationary object

If stem cells are maintained in an undifferentiated state for a long period, then they can be stored for future use, but their proliferation rate may decrease and they may undergo senescence

Engineering Contradiction:

Improveundifferentiated state maintenanceVSAvoidproliferation rate

Core Design Contradiction:

Duration of action of stationary objectVSProductivity

Solution Approach 1:

The patent optimizes media parameters including adding N-acetyl-L-cysteine (2-10 mM), ascorbic acid (0.1-1 mM), and specific growth factors to maintain high proliferation rates and prevent senescence during long-term undifferentiated storage

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent implements continuous sphere formation and regular media replacement to maintain continuous proliferation and undifferentiated state, preventing the natural decline in proliferation that occurs with simple long-term culture

Inventive Principle:

Principle #20Continuity of useful action

3Quantity of substance

If adipose tissue is used as a source of stem cells, then large amounts of cells can be obtained, but the cells are difficult to isolate and maintain in vitro

Engineering Contradiction:

Improvecell yieldVSAvoidisolation and maintenance difficulty

Core Design Contradiction:

Quantity of substanceVSEase of manufacture

Solution Approach 1:

The patent segments the adipose tissue into small pieces and uses enzymatic digestion (collagenase treatment) to release individual stem cells, enabling efficient isolation from the abundant adipose tissue while maintaining cell viability

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent optimizes isolation parameters including collagenase concentration (0.1-1%), incubation time (30-120 minutes), and filtration conditions to maximize stem cell yield while removing contaminating adipose cells and debris

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS7807461B2Multipotent stem cells derived from human adipose tissue and cellular therapeutic agents comprising the same

Publication Date: 2010.10.05 K STEMCELL CO LTD

AI summary

This invention relates to human adipose tissue-derived multipotent adult stem cells. More particularly, the invention relates to human adipose tissue-derived multipotent stem cells, which can be maintained in an undifferentiated state for a long period of time by forming spheres and have high proliferation rates, as well as methods for isolating and maintaining the adult stem cells, and methods for differentiating the multipotent adult stem cells into nerve cells, fat cells, cartilage cells, osteogenic cells and insulin-releasing pancreatic beta-cells. Also, the invention relates to cellular therapeutic agents for treating osteoarthritis, osteoporosis and diabetes and for forming breast tissue, which contain the differentiated cells or the adult stem cells. Although the multipotent stem cells are adult stem cells, they have the ability to differentiate into osteogenic cells, nerve cells, astrocytes, fat cells, chrondrogenic cells or insulin-releasing pancreatic beta-cells, and so are effective in treating osteoporosis, osteoarthritis, nerve disease, diabetes, etc. Also, the stem cells form spheres in a serum-free medium containing CORM-2, and thus can be maintained in an undifferentiated state for a long period of time. Also, the stem cells have very high proliferation rates. Accordingly, the stem cells are useful as cellular therapeutic agents.