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Updated: Jun 19, 2026
An Enzyme-free Method for Isolation and Expansion of Human Adipose-derived Mesenchymal Stem Cells
Published on: December 16, 2019
An alternative method for the isolation of mesenchymal stromal cells derived from lipoaspirate samples
Leandra S Baptista1, Ronaldo J F C do Amaral, Rosana B V Carias
1APABCAM, Clementino Fraga Filho University Hospital, Federal University of Rio de Janeiro, RJ, Brazil. leandra.baptista@gmail.com
Background Aims:
Since initial methods were developed for isolating cells from adipose tissue, little has been done to improve mesenchymal stromal cell (MSC) yield. The aim of the present study was to isolate a population of MSC from lipoaspirate samples without tissue digestion and to assess the possibility of cryopreserving the freshly isolated cells.
Methods:
A population of MSC was isolated from 13 patients' lipoaspirate samples by mechanical dissociation. Mechanically processed lipoaspirate adipose tissue (MPLA) cells were characterized after in vitro cell expansion by morphologic analysis, expression of MSC surface markers and differentiation assays.
Results:
Mechanical dissociation yielded a large quantity of adherent MSC both after standard and vibro-assisted liposuction. Preservation of lipoaspirate samples at 4 degrees C for 1 or 2 days until the mechanical procedure did not change the MPLA cell content. It was possible to store freshly isolated MPLA cells by cryopreservation without loss of the MSC population. Adherent MPLA cells were negative for CD45 and CD31 and positive for CD34, CD105, CD44 and CD90. They also showed adipogenic, osteogenic and chondrogenic potentials similar to MSC populations from other sources as already described in the literature.
Conclusions:
MSC can be isolated from human lipoaspirate samples by the mechanical procedure described in this study with a significant reduction in time and cost. Together with cryopreservation of freshly isolated MPLA cells, this has made it easier to harvest and store MSC for therapeutic applications such as soft-tissue augmentation and tissue engineering.
Insights
This study presents a new mechanical method for isolating mesenchymal stromal cells (MSCs) from lipoaspirate, reducing time and cost. Freshly isolated MSCs can be cryopreserved for future therapeutic use.
Area of Science:
- Regenerative Medicine
- Cell Biology
- Tissue Engineering
Background:
- Mesenchymal stromal cell (MSC) isolation methods from adipose tissue have seen limited improvement.
- Current techniques often involve time-consuming tissue digestion.
Purpose of the Study:
- To develop a method for isolating MSCs from lipoaspirate without tissue digestion.
- To assess the cryopreservation potential of freshly isolated MSCs.
Main Methods:
- Mechanical dissociation of lipoaspirate samples from 13 patients.
- Characterization of mechanically processed lipoaspirate (MPLA) cells via morphology, surface marker expression, and differentiation assays.
Main Results:
- Mechanical dissociation yielded abundant adherent MSCs from various liposuction techniques.
- MPLA cells maintained viability and MSC characteristics after 1-2 days of cold storage and cryopreservation.
- MPLA cells expressed typical MSC markers (CD34, CD105, CD44, CD90) and exhibited multilineage differentiation potential.
Conclusions:
- A mechanical isolation procedure offers a faster, more cost-effective method for obtaining MSCs from human lipoaspirate.
- The ability to cryopreserve freshly isolated MPLA cells simplifies MSC harvesting and storage for therapeutic applications like soft-tissue augmentation and tissue engineering.
