Mesenchymal stromal cell cryopreservation.
Abstract: The advent of stem cells and stem cell-based therapies for specific diseases requires particular knowledge of laboratory procedures, which not only guarantee the continuous production of cells, but also provide them an identity and integrity as close as possible to their origin. Their cryopreservation at temperatures below -80°C and typically below -140°C is of paramount importance. This target can be achieved by incorporating high molar concentrations of cryoprotectant mixtures that preserve cells from deleterious ice crystal formation. Usually, dimethyl sulfoxide (DMSO) and animal proteins are used as protectant reagents, but unexpected changes in stem cell fate and downstream toxicity effects have been reported, limiting their wide use in clinical settings. In scientific reviews, there are not much data regarding viability of mesenchymal stromal cells (MSCs) after the freezing/thawing process. During our routine analysis, a poor resistance to cryopreservation of these cells was observed, as well as their weak ability to replicate. This is an important point in the study of MSCs; moreover, it represents a limit for preservation and long-term storage. For this reason, MSCs isolated from equine, ovine, and rodent bone marrow and equine adipose tissue were compared using different cryopreservation solutions for this study of vitality. Our findings showed the best results regarding cell viability using a solution of fetal bovine serum with addition of 10% DMSO. In particular, we noted an increase in survival of equine bone marrow MSCs. This parameter has been evaluated by Trypan blue staining at fixed times (0, 24, and 48 hours post-thaw). This result highlights the fact that equine bone marrow MSCs are the frailest we analyzed. Therefore, it could be useful to delve further into this topic in order to improve the storage possibility for these cells and their potential use in cell-based therapies.
Publication Date: 2012-06-01 PubMed ID: 24835066DOI: 10.1089/bio.2012.0005Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Animal Science
- Animal Studies
- Biotechnology
- Bone Marrow
- Cell Culture
- Cell Proliferation
- Cell Viability
- Clinical Study
- Cryopreservation
- Disease Treatment
- Equine Health
- Equine Science
- Experimental Methods
- Freezing Technique
- In Vitro Research
- Laboratory Methods
- Mesenchymal Cells
- Stem Cells
- Veterinary Medicine
- Veterinary Research
Summary
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The research article deals with the cryopreservation of stem cells to ensure their optimal preservation for future use, with a focus on mesenchymal stromal cells (MSCs). The study observed that conventional preservation methods, often involving dimethyl sulfoxide (DMSO) and animal proteins, were yielding unsatisfactory results. Hence, the authors proceeded to study different cryopreservation solutions and their effect on cell viability.
Background
- The cryopreservation of stem cells is crucial to their lifecycle and functionality, with temperatures usually falling below -80°C and often below -140°C. This is to help maintain the integrity of the cells and preserve them for future use.
- Traditional cryopreservation techniques involve the use of DMSO and animal proteins. However, these methods have been linked to unwanted changes in the cells themselves and potential toxic effects, making them less than ideal for clinical use.
Poor Cryopreservation Results with MSCs
- The study undertaken by the authors noted that MSCs were particularly resistant to the freezing/thawing process integral to cryopreservation, and thus were also weak in their capacity for replication.
- This is a significant limit for the long-term storage and preservation of these cells, which are intern integral to stem cell-based therapies.
The Experiment
- The authors used different cryopreservation solutions to see the difference each made on the resistance to cryopreservation of MSCs from various animal sources (equine, ovine, and rodent bone marrow and equine adipose tissue).
- Therefore, the main aim of the experiment was to find a better way to freeze and preserve these cells for future use.
Results
- The results indicated that the best cryopreservation solution involved the use of fetal bovine serum along with a 10% addition of DMSO. This yielded the best results in relation to cell viability after the freezing/thawing process.
- The survival rate for equine bone marrow MSCs was particularly notable, despite being the frailer MSCs analyzed. This outcome was determined through Trypan blue staining methods at fixed post-thaw points.
Conclusion
- The conclusion drawn from these findings was that while there is still much work to be done, the successful cryopreservation of MSCs can be improved upon by altering the composition of the freezing solution.
- This will help improve the storage possibility for these cells and, consequently, their potential use in future cell-based therapies.
Cite This Article
APA
Renzi S, Lombardo T, Dotti S, Dessì SS, De Blasio P, Ferrari M.
(2012).
Mesenchymal stromal cell cryopreservation.
Biopreserv Biobank, 10(3), 276-281.
https://doi.org/10.1089/bio.2012.0005 Publication
Researcher Affiliations
- 1 Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZSLER) , Brescia, Italy .
MeSH Terms
- Adipose Tissue / cytology
- Animals
- Cell Survival
- Cells, Cultured
- Cryopreservation / methods
- Cryoprotective Agents / pharmacology
- Dimethyl Sulfoxide / pharmacology
- Horses
- Mesenchymal Stem Cells / cytology
- Organ Preservation Solutions / chemistry
- Rats
- Sheep
- Tissue Banks
Citations
This article has been cited 6 times.- Oyarzo R, Valderrama X, Valenzuela F, Bahamonde J. Bovine Fetal Mesenchymal Stem Cells Obtained From Omental Adipose Tissue and Placenta Are More Resistant to Cryoprotectant Exposure Than Those From Bone Marrow.. Front Vet Sci 2021;8:708972.
- Crippa S, Santi L, Berti M, De Ponti G, Bernardo ME. Role of ex vivo Expanded Mesenchymal Stromal Cells in Determining Hematopoietic Stem Cell Transplantation Outcome.. Front Cell Dev Biol 2021;9:663316.
- Bahsoun S, Coopman K, Akam EC. The impact of cryopreservation on bone marrow-derived mesenchymal stem cells: a systematic review.. J Transl Med 2019 Nov 29;17(1):397.
- Fu X, Yan Y, Li S, Wang J, Jiang B, Wang H, Duan Y, Tan T, Gao F, Gong D, Niu Y, Ji W, Zheng B, Si W. Vitrification of Rhesus Macaque Mesenchymal Stem Cells and the Effects on Global Gene Expression.. Stem Cells Int 2017;2017:3893691.
- Lian RL, Guo XL, Chen JS, Guo YL, Zheng JF, Chen YW. Effects of induced pluripotent stem cells-derived conditioned medium on the proliferation and anti-apoptosis of human adipose-derived stem cells.. Mol Cell Biochem 2016 Feb;413(1-2):69-85.
- Guo X, Li S, Ji Q, Lian R, Chen J. Enhanced viability and neural differential potential in poor post-thaw hADSCs by agarose multi-well dishes and spheroid culture.. Hum Cell 2015 Oct;28(4):175-89.
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