Somatic Stem Cell Isolation Using Divalent Cation Chelating Agents
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Solution Overview
Problem
The use of human embryonic stem cells for treating degenerative or inherited diseases is hindered by ethical considerations, necessitating the development of non-embryonic stem cells that can differentiate into various cell types while avoiding ethical obstacles.
Innovation Solution
Isolation and utilization of somatic stem cells, specifically SB-1 and SB-2 cells, which are CD9+, SSEA1+, SSEA4+, CD13+, or Stro1+, and can be obtained from non-embryonic sources such as bodily fluids, differentiated into all three germ layers, and expanded in vitro without losing their pluripotency, using methods involving divalent cation chelating agents like EDTA to activate and proliferate these cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If human embryonic stem cells are used for treating degenerative or inherited diseases, then therapeutic potential is improved, but ethical considerations create obstacles
Solution Approach 1:
The invention extracts the desired therapeutic function of stem cells (pluripotency and differentiation capability) from their traditional embryonic source, and relocates it to a non-embryonic source (somatic cells). This separates the beneficial therapeutic effects from the ethical problems associated with embryo destruction, thereby resolving the contradiction between therapeutic potential and ethical considerations
Solution Approach 2:
The invention creates a copy of the embryonic stem cell state and function within somatic cells through reprogramming. The somatic cells are transformed to exhibit embryonic-like pluripotency without actually using embryos, thus copying the therapeutic functionality while avoiding the ethical issues of embryonic cell usage
2Quantity of substance
If somatic stem cells are isolated and expanded in vitro, then cell availability for treatment is improved, but maintaining pluripotency without differentiation becomes difficult
Solution Approach 1:
The invention changes the physical and chemical parameters of the culture environment, specifically using divalent cation chelating agents (like EDTA or EGTA) to alter ion concentrations. This parameter change prevents spontaneous differentiation while allowing cell expansion, thereby maintaining pluripotency stability during in vitro cultivation
Solution Approach 2:
The invention introduces divalent cation chelating agents as intermediary substances that mediate between the somatic cells and the culture environment. These agents bind divalent cations to create a specific ionic environment that suppresses differentiation signals while permitting cell proliferation, thus acting as a mediator to maintain pluripotency during expansion
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
These somatic stem cells can be used to treat degenerative disorders, muscle injuries, and muscle-degenerative diseases by administering them to subjects, offering a non-embryonic alternative for regenerative medicine, with the ability to form all somatic cells and reproductive cells, and induce differentiation into specific cell types, thus bypassing ethical concerns.
Implementation Method 1
incubating the sample with a divalent cation chelating agent (e.g., EDTA, EGTA, and sodium citrate) or heparin in a container until the sample is separated into an upper layer and a lower layer
Implementation Method 2
The isolation of SB cells from the upper layer can be conducted by methods based on cell sizes (e.g., centrifuging and filtering)
Implementation Method 3
incubating it with ADP so as to allow platelets/microparticles to precipitate for further enrichment of the stem cells
Implementation Method 4
incubating the upper layer obtained from a heparin-treated sample with a divalent cation chelating agent to activate the cell cycle of the stem cells from G0 into G1
Data Source
AI summary
A population of somatic stem cells and a method of preparing same. Also disclosed are two subpopulations thereof and their various uses.