Cooling Muscle Tissue for High-Purity Cell Isolation
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Solution Overview
Problem
Current methods for isolating skeletal muscle-derived cells (SMDC) are time-consuming and inefficient, leading to low purity and viability, which hampers their effectiveness in treating muscle dysfunctions like urinary and anal incontinence.
Innovation Solution
A method involving cooling of muscle tissue samples, followed by enzymatic processing and resuspension in serum-containing medium, eliminates the need for pre-plating steps, resulting in highly pure and viable SMDC with enhanced myogenic and neuronal precursor cell markers, such as CD56 and A2B5, for improved neuromuscular support.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional isolation methods are used for skeletal muscle-derived cells, then the cells can be obtained, but the process is time-consuming and results in low purity and viability
Solution Approach 1:
The invention changes the physical parameter of temperature by cooling the muscle tissue sample to between -80°C and 0°C before enzymatic processing. This parameter change fundamentally alters the isolation process, enabling direct plating without pre-plating steps while achieving high purity (≥90% myogenic cells) and viability, thereby resolving the contradiction between purity and time
Solution Approach 2:
The cooling step is performed as a preliminary action before enzymatic processing and cell isolation. This pre-cooling of the tissue sample prepares the cells for subsequent steps, preserving their viability and enabling direct plating, thus eliminating time-consuming intermediate purification steps
2Productivity
If conventional isolation methods are used for skeletal muscle-derived cells, then the cells can be obtained, but the process is complex and inefficient
Solution Approach 1:
The invention segments the isolation process into distinct temperature-controlled phases: initial cooling of tissue, enzymatic digestion at controlled temperature, and direct plating. This segmentation simplifies the overall process by eliminating the need for multiple pre-plating and purification steps, thereby improving productivity while reducing complexity
Solution Approach 2:
The invention introduces temperature control as an intermediary mechanism that mediates between the tissue sample and the isolation process. By maintaining specific temperature ranges during cooling and enzymatic processing, the method simplifies subsequent steps and improves efficiency without adding complex equipment
3Manufacturing precision
If pre-plating steps are used to purify skeletal muscle-derived cells, then cell purity may improve, but the process becomes time-consuming and reduces cell viability
Solution Approach 1:
The invention changes the temperature parameter to between -80°C and 0°C before isolation, which preserves cell viability while enabling direct plating. This parameter change eliminates the need for multiple pre-plating steps that would otherwise be required to achieve high purity, thus maintaining both purity and reliability simultaneously
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 yields SMDC with high purity and viability, effectively supporting neuromuscular connections and regenerating muscle weakness, particularly in conditions like urinary and anal incontinence, by enhancing myogenic capacity and neuro-muscular support.
Implementation Method 1
cooling of a sample obtained from skeletal muscle tissue in a buffer
Implementation Method 2
processing and cooling of the sample; resuspending the sample of step (b) in medium with serum comprising at least one enzyme
Data Source
AI summary
The present invention relates to methods for obtaining skeletal muscle derived cells (SMDC), and the use of SMDCs in a method of preventing and/or treating neuromyopathies and/or myopathies, wherein the neuromyopathy and/or myopathy is incontinence, in particular a urinary and/or an anal or fecal incontinence.