Mammalian Tissue Perfusion for Disinfected Cell Suspensions
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Solution Overview
Problem
Conventional decontamination methods for sensitive tissues like liver cells, such as heating, autoclaving, or irradiation, are ineffective in maintaining cell viability, and antibiotic-based processes often fail to ensure microbial sterility, especially when contamination is internal and difficult to access.
Innovation Solution
A process involving perfusion of mammalian tissues with an antibiotic composition, followed by enzymatic treatment using collagenase and proteinase, to obtain disinfected single-cell suspensions while maintaining cell viability and ensuring thorough disinfection, including hard-to-reach areas within the tissue.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional decontamination methods (heating, autoclaving, irradiation) are used, then microbial sterility is improved, but cell viability is worsened
Solution Approach 1:
The patent replaces mechanical/thermal decontamination methods (autoclaving, heating, irradiation) with a chemical approach using antibiotic compositions. The antibiotics are introduced via perfusion through the tissue's vascular system, allowing decontamination without the harmful physical effects that damage cell viability.
Solution Approach 2:
The patent uses an intermediary substance (antibiotic composition) that mediates between the decontamination goal and the preservation of cell viability. The antibiotics serve as a chemical intermediary that kills microorganisms without the destructive physical effects of conventional methods, thus protecting cell integrity.
2Reliability
If antibiotic-based decontamination processes are used, then microbial sterility is improved, but disinfection completeness is worsened (surface only)
Solution Approach 1:
The patent employs a hydraulic approach by using perfusion through the tissue's vascular system. The antibiotic-containing fluid is pumped through blood vessels, allowing the antibiotics to be transported deep into the tissue interior and reach hard-to-access areas, achieving comprehensive disinfection rather than surface-level treatment.
Solution Approach 2:
The patent transitions from surface-level decontamination to three-dimensional penetration into the tissue interior. By utilizing the vascular network's distribution system, the antibiotic composition reaches microorganisms throughout the entire tissue volume, including deep and hidden areas, achieving complete disinfection.
3Reliability
If long exposure times to antibiotics are used, then microbial sterility is improved, but tissue viability is worsened
Solution Approach 1:
The patent optimizes the concentration parameters of the antibiotic composition and adjusts perfusion flow rates to achieve effective disinfection within shorter exposure times. By controlling these parameters, the system achieves microbial sterility without requiring prolonged exposure that would harm tissue viability.
4Reliability
If antibiotics are used for decontamination, then microbial sterility is improved, but compatibility with sterility verification system is worsened
Solution Approach 1:
The patent segments the decontamination process into distinct phases: first, perfusion with antibiotic composition for disinfection; second, washing steps to remove residual antibiotics; third, sterility verification. This segmentation allows the antibiotic treatment to occur without interfering with subsequent sterility testing, as the antibiotics are removed before verification.
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 achieves effective and rapid disinfection of liver cells, maintaining their viability and sterility, with over 65% of liver cell preparations being disinfected, and allows for accurate sterility testing, overcoming the limitations of surface-only disinfection and cell damage in previous methods.
Implementation Method 1
perfusing mammalian tissue with a liquid antibiotic composition... achieving effective and rapid disinfection of liver cells
Implementation Method 2
carrying out an enzymatic treatment of the tissue, in particular to obtain a single-cell suspension... using collagenase and proteinase
Data Source
AI summary
In a process for preparing disinfected preparations from mammalian tissues, mammalian tissue is perfused at one step with an enzyme-free liquid antibiotic composition that includes at least one antibiotic and is contained in at least one perfusion buffer. In a step occurring after such one step, enzymatic antibiotic-free treatment of the tissue is carried out to obtain a single-cell suspension. In another step disinfected preparations are obtained.