Stem Cell Isolation from Liposuction Aspirate via Density Gradient
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Solution Overview
Problem
Current methods for isolating mesenchymal stem cells from bone marrow are costly, labor-intensive, and painful for donors, with limited availability and difficulty in culturing these cells without inducing differentiation, while existing methods for obtaining stem cells from fat tissue require enzyme treatment and are not efficient for large-scale production.
Innovation Solution
A method involving centrifugation of liposuction aspirate to separate a cell fraction by specific gravity, using saline or Ringer's solution, and further processing with Ficoll or Percoll density gradients to collect a cell layer with lower specific gravity than erythrocytes, allowing for the isolation of stem cells without enzyme treatment and in a more efficient and large-scale manner.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If bone marrow is used as the cell source for isolating mesenchymal stem cells, then stem cells can be obtained, but the process is costly, labor-intensive, and painful for donors with limited availability
Solution Approach 1:
The invention extracts and utilizes stem cells from adipose tissue (fat) instead of bone marrow. The method involves aspirating fat tissue, treating it with collagenase to release stem cells from the extracellular matrix, and then isolating the stem cells through centrifugation and filtration. This extraction approach from an alternative source (adipose tissue) resolves the contradiction by providing abundant stem cells without the limitations of bone marrow harvesting.
Solution Approach 2:
The invention changes the source parameter from bone marrow to adipose tissue, and modifies the isolation parameters by using collagenase treatment followed by centrifugation at specific gravity conditions. This parameter change enables large-scale production of stem cells with high purity, overcoming the availability and complexity limitations of bone marrow-based methods.
2Productivity
If existing methods are used to prepare stem cells from fat tissue, then stem cells can be obtained, but enzyme treatment is required which complicates the process and reduces efficiency
Solution Approach 1:
The invention applies collagenase treatment to the aspirated fat tissue before centrifugation to pre-digest the extracellular matrix and release stem cells. This preliminary action simplifies subsequent isolation steps by already separating the stem cells from the tissue matrix, thereby improving productivity while managing complexity through a standardized enzymatic treatment protocol.
Solution Approach 2:
The invention replaces complex mechanical dissection and manual isolation methods with a standardized enzymatic digestion process followed by centrifugation. This substitution of mechanical systems with biochemical processes simplifies the overall method, improves reproducibility, and enhances productivity by enabling large-scale processing of adipose tissue.
3Reliability
If mesenchymal stem cells are isolated from bone marrow, then stem cells can be obtained, but it is difficult to culture these cells without inducing differentiation
Solution Approach 1:
The invention isolates stem cells from adipose tissue using a method that preserves their undifferentiated state and self-renewal capacity. The collagenase treatment and gentle centrifugation process maintains cell integrity and functionality, enabling the isolated stem cells to be cultured without immediate differentiation. This self-preserving isolation method resolves the contradiction by providing both high purity and ease of culture maintenance.
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 enables the simple and efficient preparation of homogeneous stem cells and precursor cells from liposuction aspirate, overcoming the limitations of existing techniques by providing a large amount of stem cells with high purity and facilitating their use in regenerative medicine.
Implementation Method 1
B) subjecting the aspirate from liposuction to centrifugation to obtain a cell fraction
Implementation Method 2
C) subjecting the cell fraction to centrifugation by specific gravity
Implementation Method 3
C) subjecting the cell fraction to centrifugation by specific gravity
Data Source
AI summary
The present invention provides a method for preparing homogeneous stem cells and/or precursor cells in a large amount, in a simple and efficient manner, from human (in particular, the subject per se). The present invention further provides a method for preparing tissue implant or graft in a large amount by the use of the stem cells and/or precursor cells of the present invention. The present inventors have unexpectedly discovered that an aspirate from liposuction contains a large number of stem cells and have established a method for preparing stem cells from such an aspirate from liposuction, and thereby achieved the above-mentioned object.