Umbilical Cord Vessel MSC Isolation via Segmented Collagenase Incubation
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Solution Overview
Problem
Current methods for obtaining mesenchymal stem cells (MSCs) from the perivascular space of the umbilical cord are time-consuming, result in low cell yield, and have safety concerns due to prolonged enzyme treatment, leading to contamination and reduced proliferative activity, making them unsuitable for mass therapeutic use.
Innovation Solution
A method involving two incubation cycles with a collagenase solution in the umbilical cord vessel, followed by washing and centrifugation, to increase the proportion of MSCs, reduce processing time, and enhance culture purity and safety, by selectively isolating different cell populations and minimizing collagenase exposure.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If prolonged enzyme treatment (24 hours) is used to isolate MSCs from perivascular space, then cell isolation can be achieved, but cell viability decreases and processing time increases
Solution Approach 1:
The patent divides the enzyme treatment process into two separate incubation cycles with collagenase solution, each lasting 15-45 minutes. This segmentation allows selective release of different cell populations (first cycle releases endothelial cells, second cycle releases MSCs) while limiting total enzyme exposure time to 30-90 minutes, thereby maintaining cell viability and reducing processing time compared to continuous 24-hour treatment
Solution Approach 2:
The patent employs periodic action by using two distinct incubation cycles with collagenase solution separated by washing steps. The first incubation cycle (15-45 minutes) targets endothelial cells, followed by washing and removal of collagenase. The second incubation cycle (25-65 minutes) then targets MSCs. This periodic application of enzyme treatment minimizes cumulative enzyme exposure while achieving complete cell isolation, resolving the contradiction between treatment effectiveness and cell viability
2Productivity
If single incubation cycle with collagenase is used, then processing time is reduced, but cell yield and purity are insufficient
Solution Approach 1:
The patent segments the cell isolation process into two sequential incubation cycles. The first cycle (15-45 minutes) selectively releases endothelial cells from the perivascular space. After washing and removing collagenase, the second cycle (25-65 minutes) selectively releases MSCs. This segmentation enables complete cell yield by targeting different cell populations in separate steps, achieving both high productivity and adequate processing time
Solution Approach 2:
The patent applies partial action by using two separate incubation cycles instead of one prolonged cycle. Each cycle uses optimized time parameters (15-45 minutes for first cycle, 25-65 minutes for second cycle) that are sufficient for complete cell release but limit total enzyme exposure. This partial application of enzyme action at optimized intervals achieves complete cell yield while controlling processing time
3Reliability
If prolonged enzyme treatment is used, then complete cell isolation is achieved, but contamination and safety issues arise
Solution Approach 1:
The patent segments the enzyme treatment into two distinct cycles with washing steps in between. The first cycle (15-45 minutes) isolates endothelial cells, followed by complete washing and collagenase removal. The second cycle (25-65 minutes) then isolates MSCs. This segmentation minimizes cumulative collagenase exposure time (totaling 30-90 minutes vs. 24 hours), reducing contamination risk while maintaining complete cell isolation and safety
Solution Approach 2:
The patent rushes through the enzyme treatment process by using two short incubation cycles (15-45 minutes and 25-65 minutes) instead of one prolonged 24-hour cycle. The intermediate washing steps remove collagenase between cycles, preventing continuous enzyme exposure. This rushing approach achieves complete cell isolation in 30-90 minutes total time, significantly reducing contamination risk and improving cell safety while maintaining processing completeness
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 significantly reduces the time required to obtain MSC cultures, increases cell yield and purity, and maintains the viability and proliferation potential of MSCs, making them more suitable for regenerative therapies.
Implementation Method 1
filling the umbilical cord vessel with 0.01-0.5% collagenase solution; incubating the umbilical cord at 37° C. for 15-45 minutes
Implementation Method 2
The first and second balanced salt solution are centrifugated to obtained precipitating pellets
Data Source
AI summary
The method of the present invention includes cutting out a umbilical cord segment; filling the vessel of the umbilical cord segment with 0.01-0.5% collagenase solution; sealing both ends of the umbilical cord segment and maintaining it at a temperature of 37° C. for at least 15-45 minutes; wash the vessel with balanced normal saline solution; fill the vessel with a new 0.01-0.5% collagenase solution; seal both ends of the umbilical cord and incubate at 37° C. for at least 25-65 minutes; and equilibrate The vessel is washed with saline solution, the washed liquid is centrifuged, and the resulting pellet is incubated in a selective environment. The present invention allows to increase the yield and purity of obtained mesenchymal stem cell cultures and in some cases can reduce the time required to obtain mesenchymal stem cells.