Equine Amniotic Fluid Stem Cell Isolation for Homogeneous Populations
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Solution Overview
Problem
Current methods for isolating and culturing equine stem cells face challenges due to invasive procedures, limited availability, and heterogeneity issues, particularly in obtaining homogeneous adult stem cells for therapeutic use in racehorses, where the known markers for adult stem cells are not well-developed and the isolation process is costly and inefficient.
Innovation Solution
The isolation and characterization of equine amniotic fluid-derived multipotent stem cells (eAF-MSCs) using centrifugation and specific culture media like EGM-2 and DMEM, which exhibit distinct immunological characteristics and maintain an undifferentiated state for 14 passages or more, allowing for the production of homogeneous stem cell populations with enhanced self-renewal and differentiation capabilities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If stem cells are isolated from adult tissues such as bone marrow or adipose tissue, then stem cells can be obtained for therapeutic use, but the isolation procedure becomes invasive and difficult to perform
Solution Approach 1:
The patent extracts stem cells from amniotic fluid, a non-invasive source obtained during foaling, rather than from adult tissues like bone marrow or adipose tissue. This extraction from an alternative, more accessible source resolves the contradiction between obtaining stem cells and avoiding invasive procedures
Solution Approach 2:
The patent utilizes amniotic fluid collected during the natural foaling process as a preliminary source of stem cells. By preparing and storing this fluid beforehand, the need for invasive procedures at the time of cell collection is eliminated, resolving the contradiction between stem cell acquisition and procedural ease
2Quantity of substance
If stem cells are cultured from adult tissues, then stem cells can be obtained, but the cells show heterogeneity and difficulty in maintaining undifferentiated state
Solution Approach 1:
The patent changes the source parameter from adult tissues to amniotic fluid-derived cells, and optimizes culture conditions including specific growth factors and media composition. These parameter changes enable maintenance of undifferentiated state and produce homogeneous stem cell populations, resolving the contradiction between cell quantity and composition stability
3Manufacturing precision
If stem cells are isolated using cell sorting with antibodies, then homogeneous stem cell populations can be obtained, but the cost increases and the process becomes complex
Solution Approach 1:
The patent extracts and utilizes specific growth factors and signaling molecules from amniotic fluid that naturally promote stem cell proliferation and maintain homogeneity. This extraction-based approach replaces complex antibody sorting devices with a simpler biological extraction method, resolving the contradiction between manufacturing precision and device complexity
4Quantity of substance
If large number of stem cells are required for racehorse therapy, then therapeutic effectiveness can be improved, but obtaining sufficient cells becomes difficult from adult tissues
Solution Approach 1:
The patent collects and stores amniotic fluid during the natural foaling process as a preliminary cell source. This preliminary collection provides a abundant supply of stem cells without requiring invasive procedures later, resolving the contradiction between obtaining large cell numbers and maintaining productivity
Solution Approach 2:
The patent changes the source parameter to amniotic fluid, which naturally contains higher concentrations of stem cells compared to adult tissues. Combined with optimized culture conditions, this parameter change enables high-yield cell production, resolving the contradiction between cell quantity and production efficiency
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 provides a reliable and efficient source of homogeneous equine adult stem cells with superior self-renewal and differentiation abilities, suitable for large-scale production and therapeutic applications, particularly for treating injuries and diseases in racehorses, such as osteoarthritis and bone loss.
Implementation Method 1
isolating and characterizing equine amniotic fluid-derived multipotent stem cells
Data Source
Figure 1A~1C
Figure 2
Figure 3A~3B
AI summary
The present invention relates to equine amniotic fluid-derived multipotent stem cells (eAF-MSCs) and a preparation method thereof. More particularly, the present invention relates to equine amniotic fluid-derived multipotent stem cells which exhibit all negative immunological characteristics with respect to the human markers, CD19, CD20, CD28, CD31, CD34, CD38, CD41a, CD62L, CD62P and CD200, and exhibit all positive immunological characteristics with respect to the human markers, CD44, CD90 and CD105, and have the ability to differentiate into ectoderm, mesoderm or endoderm-derived cells. The present invention confirmed that equine amniotic fluid can be a source of equine adult multipotent stem cell supply, and the equine amniotic fluid-derived multipotent stem cells exhibit excellent self-renewal ability and differentiation capability. Therefore, the cells can be used as an active ingredient for cell therapy of horses, which requires a large amount of cells. In particular, the stem cells of the present invention having excellent self-renewal ability and differentiation capability are useful for the treatment of bone, tendon, or muscle injuries and bone loss diseases of racehorses.