Mesenchymal Stem Cell Extraction from Wharton's Jelly via Red Blood Cell Lysis
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Solution Overview
Problem
The extraction of mesenchymal stem cells from umbilical cord tissue is hindered by interference from red blood cells, which affects cell viability and purity, limiting their quantity and quality for clinical applications.
Innovation Solution
The method involves using a red blood cell lysis buffer and a serum-free culture system to separate and culture mesenchymal stem cells from Wharton's jelly tissue, specifically treating the tissue with a buffer containing NH4Cl and Na2-EDTA, followed by serum-free medium with β-mercaptoethanol, non-essential amino acids, and recombinant human basic fibroblast growth factor, to enhance cell separation and purity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If enzyme digestion and tissue adherence method are used to extract mesenchymal stem cells from umbilical cord, then the extraction process is simple, but the presence of red blood cells interferes with cell growth and reduces stem cell viability
Solution Approach 1:
The patent applies preliminary action by performing red blood cell lysis treatment before the main culture process. The Wharton's jelly tissue is treated with red blood cell lysis buffer to remove red blood cells prior to enzymatic digestion and culture, preventing their interference with stem cell growth and improving viability without complicating the overall process
Solution Approach 2:
The patent converts the harmful presence of red blood cells into a beneficial separation process. By using red blood cell lysis buffer, the harmful interference from red blood cells is transformed into a useful purification step that enhances stem cell viability while maintaining process simplicity
2Productivity
If traditional culture medium with serum is used, then cell growth is supported, but the purity and quality of mesenchymal stem cells are compromised due to red blood cell interference
Solution Approach 1:
The patent applies the extraction principle by removing red blood cells from the culture system through lysis buffer treatment. This selective removal of the harmful component (red blood cells) allows the beneficial component (mesenchymal stem cells) to grow with higher purity and quality, resolving the contradiction between growth support and purity maintenance
3Quantity of substance
If red blood cells are present in the culture, then the initial cell population is abundant, but the growth space is limited and cell viability is reduced
Solution Approach 1:
The patent converts the harmful occupation of growth space by red blood cells into a beneficial purification opportunity. By applying red blood cell lysis buffer, the space occupied by harmful red blood cells is freed up, creating more growth space for mesenchymal stem cells while maintaining abundant initial cell population
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 approach effectively mitigates the interference from red blood cells, improving the purity and viability of mesenchymal stem cells, enabling their successful extraction and storage for clinical use.
Implementation Method 1
adding red blood cell lysis buffer with a volume of 1-3 times the volume of the tissue blocks, and treating the tissue blocks with the buffer at room temperature for 2-5 minutes
Data Source
AI summary
Provided is a method for separating and extracting mesenchymal stem cells from the human umbilical cord. The method uses healthy neonatal umbilical cord tissue; after cleaning and disinfection, mechanically pulverising same, separating the Wharton's jelly, and after treating with erythrocyte lysate, carrying out suspension culture in a serum-free culture medium. Replacing the liquid every 3-5 days; after the plate adherence rate reaches 30-70%, carrying out trypsin digestion, and then collecting the cells by centrifugation for passage amplification, until the rate of confluence of the cells reaches 80-90% confluence, thereby obtaining high purity umbilical cord mesenchymal stem cells.