Equine Amniotic Membrane Stem Cells Non-Invasive Isolation
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Solution Overview
Problem
Current methods for isolating and culturing stem cells from equine tissues, such as adipose and bone marrow, are invasive, inefficient, and result in heterogeneous cell populations, limiting their therapeutic application due to the lack of known surface markers and difficulty in maintaining undifferentiated states for extended periods.
Innovation Solution
Isolation and culture of equine amniotic membrane-derived mesenchymal stem cells (eAM-MSCs) using low-glucose Dulbecco's modified Eagle medium, which exhibit negative responses to human markers CD19, CD20, CD28, CD31, CD34, CD38, CD41a, CD62L, and CD200, and positive responses to CD44, CD90, and CD105, allowing for prolonged undifferentiated maintenance and multilineage differentiation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If stem cells are isolated from equine adipose or bone marrow tissues, then stem cells can be obtained, but the isolation methods are invasive and result in heterogeneous cell populations
Solution Approach 1:
The patent extracts stem cells from the amniotic membrane, a non-invasive tissue easily accessible during routine equine Cesarean sections. This eliminates the need for invasive bone marrow aspirations or adipose surgeries while obtaining sufficient stem cell material for therapeutic applications.
Solution Approach 2:
The amniotic membrane is typically discarded as waste after equine Cesarean sections. The patent recovers stem cells from this discarded tissue, transforming it from useless waste into a valuable source of therapeutic stem cells without requiring additional invasive procedures.
2Productivity
If stem cells are cultured from equine tissues, then cell populations can be expanded, but the cells differentiate prematurely and cannot be maintained in undifferentiated state for extended periods
Solution Approach 1:
The patent employs specific culture conditions including low-glucose DMEM medium, 5% CO2 atmosphere, and 37°C temperature to maintain stem cell undifferentiated state. These parameter optimizations allow extensive cell passage (14+ passages) while preserving stemness and preventing premature differentiation.
3Quantity of substance
If conventional stem cell isolation methods are used, then stem cells can be obtained, but the lack of known surface markers makes characterization difficult
Solution Approach 1:
The patent replaces the need for complex surface marker profiling with functional characterization methods. Instead of relying on known surface markers for identification, the patent uses differentiation potential assessment and immunophenotyping with antibodies against CD markers to characterize eAM-MSCs, simplifying the characterization process.
4Adaptability or versatility
If equine stem cells are used for therapy, then treatment options expand, but the heterogeneous cell populations limit therapeutic efficacy
Solution Approach 1:
The patent applies local quality control by selecting and expanding only the stem cell population with specific characteristics (CD44+, CD90+, CD105+) from the amniotic membrane tissue. This ensures therapeutic preparations contain homogeneous, therapeutically relevant cells rather than mixed heterogeneous populations.
Data Source
AI summary
The present invention relates to equine amniotic membrane-derived mesenchymal stem cells (eAM-MSCs) and a preparation method thereof. More specifically, the present invention relates to equine amniotic membrane-derived mesenchymal stem cells, which show negative immunological responses to all of the human markers CD19, CD20, CD28, CD31, CD34, CD38, CD41a, CD62L, CD62P and CD200, and positive immunological responses to all of the human markers CD44, CD90 and CD105, and have the ability to be maintained in an undifferentiated state for 14 passages or more and the ability to differentiate into ectoderm, mesoderm and endoderm-derived cells.