Cardiac Tissue-Derived Pluripotent Stem Cell Isolation
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Solution Overview
Problem
Current stem cell therapies for cardiac tissue regeneration face challenges due to the lack of clinically applicable pluripotent stem cells with high differentiation capability into cardiac myocytes, poor proliferative ability, and difficulty in isolation and large-scale culture of existing myocardial stem cells.
Innovation Solution
A method involving enzymatic treatment of cardiac tissue fragments to create a cell suspension, followed by density gradient separation and suspension culture with fibroblast growth factor and epidermal growth factor to select and separate cells forming floating spheres, resulting in c-kit-negative, CD31-negative, CD34-negative, CD105-positive pluripotent stem cells capable of differentiating into cardiac myocytes and other cell types.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If cardiac tissue-derived stem cells are used for myocardial regeneration, then differentiation capability into cardiac myocyte is improved, but proliferative ability deteriorates
Solution Approach 1:
The patent applies parameter changes by modifying culture conditions including specific growth factors (basic fibroblast growth factor at 10-100 ng/mL, epidermal growth factor at 10-100 ng/mL), serum concentration (10-20% fetal bovine serum), and culture medium composition to simultaneously enhance both differentiation capability and proliferative ability of cardiac tissue-derived stem cells
2Adaptability or versatility
If myocardial stem cells are isolated from cardiac tissue, then cell source is improved, but isolation difficulty and culture complexity increase
Solution Approach 1:
The patent applies segmentation by dividing the isolation and culture process into distinct sequential steps: (1) enzymatic digestion of cardiac tissue with collagenase and trypsin, (2) density gradient centrifugation using Percoll to separate stem cells, (3) suspension culture in specific growth medium, and (4) selection of adherent cells. This systematic segmentation simplifies the overall complex process of obtaining pure cardiac tissue-derived stem cells
Solution Approach 2:
The patent uses Percoll density gradient centrifugation as an intermediary technique to separate cardiac tissue-derived stem cells from other cardiac cells based on density differences. This intermediary method enables efficient isolation without requiring complex purification steps
3Reliability
If existing myocardial stem cells are used for transplantation, then therapeutic potential is improved, but clinical applicability deteriorates due to poor proliferative ability
Solution Approach 1:
The patent modifies culture parameters including adding basic fibroblast growth factor (10-100 ng/mL) and epidermal growth factor (10-100 ng/mL) to the culture medium, maintaining 10-20% fetal bovine serum concentration, and controlling culture at 37°C with 5% CO2 to enhance proliferative ability while maintaining therapeutic potential for clinical transplantation
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
The method produces pluripotent stem cells with excellent differentiation and self-renewal capabilities, making them suitable for regenerative therapy, particularly for cardiac diseases, offering a novel alternative to heart transplantation.
Implementation Method 1
separating a group of cardiac tissue-derived cells from the cell suspension by the density gradient method
Data Source
AI summary
An object of the present invention is to provide a stem cell applicable to regenerative therapeutic method, and to provide a technique to carry out regenerative therapy using the cell. A collected cardiac tissue fragment is enzymatically treated to prepare a cell suspension. Then using the cell suspension, following steps are carried out: (1) separation of cells by the density gradient method, (2) suspension-culture in a culture medium containing fibroblast growth factor and epidermal growth factor and (3) selection and separation of cells forming a floating sphere to obtain pluripotent stem cells. Thus-obtained pluripotent stem cells are used to carry out regenerative therapy.