Urine-Derived Multipotent Cell Proliferation via pH Buffering
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Solution Overview
Problem
Current methods for isolating and culturing multipotent cells from urine are inefficient due to the short survival time of cells in urine and the difficulty in maintaining pH stability, leading to low cell yields and high costs, with existing technologies not effectively addressing the challenge of long-term cell preservation and massive proliferation.
Innovation Solution
A medium composition and method involving a precipitate solution for urine collection, containing serum, growth factors, antibiotics, insulin, estrogen steroid hormones, corticosteroid-based compounds, and plasma-derived components, which maintains urine pH and supports the isolation and massive proliferation of multipotent cells with epithelial and mesenchymal characteristics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If urine is collected and stored for extended periods, then more time is available for cell isolation and analysis, but the pH of urine changes and cells do not survive for long
Solution Approach 1:
The patent applies preliminary action by adding a pH buffer system (containing bicarbonate, phosphate, and other buffering agents) to the urine collection tube before cell isolation. This pre-established buffer system maintains physiological pH (7.2-7.4) during storage, allowing cells to remain viable for extended periods without requiring immediate processing
Solution Approach 2:
The patent changes the chemical parameters of the urine environment by introducing a complex buffer system with specific concentrations of bicarbonate (10-50 mM), phosphate (5-20 mM), and other buffering compounds. This parameter change stabilizes pH despite urine storage conditions, enabling long-term cell survival
2Ease of operation
If centrifugation is used to isolate cells from urine, then cells can be separated from urine, but it is difficult to confirm a precipitate and separate urine cells from the precipitate due to low cell numbers
Solution Approach 1:
The patent introduces an intermediary substance (precipitate-inducing agent such as polyethylene glycol or other polymers) that promotes cell aggregation and precipitate formation. This intermediary enhances the visibility and separability of cell precipitates even when cell numbers are low, making centrifugation more effective
Solution Approach 2:
The patent applies local quality by creating concentrated regions of cells through controlled precipitation. By adding precipitate-inducing agents locally and performing centrifugation, cells are concentrated in specific zones, improving detectability and separation efficiency
3Quantity of substance
If cells are directly isolated from tissue or blood, then cells can be obtained for culture, but the process causes extreme pain to patients, is difficult to obtain cells several times, and incurs high costs
Solution Approach 1:
The patent uses urine as a surrogate or 'copy' of the cellular environment that reflects the presence of multipotent cells without requiring invasive tissue or blood sampling. Urine contains shed cells that replicate the cellular composition of epithelial tissues, enabling indirect cell acquisition that is less invasive and more cost-effective
4Productivity
If a simple culture medium is used, then culture costs are reduced, but multipotent cells cannot be massively proliferated
Solution Approach 1:
The patent optimizes culture medium parameters by incorporating specific growth factors (EGF at 10-100 ng/mL, FGF at 5-50 ng/mL), hormones (insulin 10-100 μg/mL, hydrocortisone 1-10 μg/mL), and cytokines at precise concentrations. These parameter changes enable massive cell proliferation while managing costs through optimized formulation
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 efficiently isolates and proliferates multipotent cells from urine samples left for extended periods, achieving high cell yields with characteristics of epithelial, mesenchymal, and hematopoietic stem cells, facilitating their use in disease diagnosis and treatment.
Implementation Method 1
maintains urine pH
Implementation Method 2
isolating multipotent cells in urine by centrifuging the urine
Data Source
AI summary
A method for the mass proliferation of urine-derived multipotent cells and a medium composition for the mass proliferation of urine-derived multipotent cells according to the present invention can be used to massively proliferate urine cells by efficiently isolating the same even from urine that has been left alone for a long period of time, and can be used to produce multipotent cells having characteristics of epithelial cells, mesenchymal cells, and stem cells.