Conditioned medium: a new alternative for cryopreservation of equine umbilical cord mesenchymal stem cells.
Abstract: Cryopreservation is a feasible alternative to maintaining several cell lines, particularly for immediate therapeutic use, transportation of samples, and implementation of new in vitro studies. This work parts from the hypothesis that the medium of cryopreservation composed by 90% of conditioned medium (CM) supports cryopreservation of equine umbilical cord intervascular matrix mesenchymal stem cells (UCIM-MSCs), allowing the maintenance of the biological properties for the establishment of cell banks intended for therapeutic use and in vitro studies. Thus, we evaluated the viability, apoptosis/necrosis rates, immunophenotypic profile (IP), chromosomal stability, clonicity, and differentiation potential of UCIM-MSCs cryopreserved with four different mediums (with FBS: M1, M3, M4 and without FBS: M2). After 3 months of cryopreservation, samples were thawed and analyzed. The potential of differentiation in the mesodermal lineages, clonicity, and the chromosomal stability were maintained after cryopreservation of UCIM-MSCs with medium containing FBS. Changes (P < 0.05) at IP for some markers were observed at cells cryopreserved with medium M1-M3. Only the UCIM-MSCs cryopreserved with the CM (M4) had similar viability post-thaw (P = 0.23) when compared with fresh cells. We proved the hypothesis that the medium of cryopreservation containing CM supports the cryopreservation of UCIM-MSCs, at the experimental conditions, being the medium that better maintains the biological characteristics observed at fresh cells. Thus, future studies of UCIM-MSCs secretome should be conducted to better understand the beneficial and protective effects of the CM during the freezing process.
© 2017 International Federation for Cell Biology.
Publication Date: 2017-02-02 PubMed ID: 27888544DOI: 10.1002/cbin.10708Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The study proposes the usage of conditioned medium (CM) as an efficient medium for the cryopreservation of horse umbilical cord mesenchymal stem cells. This method ensures the cells maintain their original biological properties and the capacity for therapeutic use and cell culture studies after defrosting.
Objective and Hypothesis
- The research is inspired by the hypothesis that a cryopreservation medium made up of 90% conditioned medium (CM) can effectively help in the cryopreservation of equine umbilical cord intervascular matrix mesenchymal stem cells (UCIM-MSCs), preserving their biological attributes, so they can be used for creation of cell banks for therapeutic applications and in vitro studies.
Methods
- Researchers analyzed the viability, apoptosis/necrosis rates, immunophenotypic profile (IP), chromosomal stability, clonicity (replication of identical cells), and the potential of UCIM-MSCs to differentiate into different cell types after cryopreservation using four distinct mediums. These mediums are: M1, M3, M4 (all containing FBS or Fetal Bovine Serum), and M2 (without FBS).
- After 3 months of cryopreservation, the samples were thawed and examined to determine the efficacy of each medium.
Findings
- UCIM-MSCs that were cryopreserved with media containing FBS (M1, M3, and M4), retained their differentiation potential into mesodermal lineages, chromosomal stability, and clonicity. On the other hand, some changes (P < 0.05) in the immunophenotypic profile were observed in cells cryopreserved with the M1-M3 media.
- Cells cryopreserved using medium M4 (CM) demonstrated similar viability to fresh cells post-thaw, suggesting that CM is highly effective for cryopreservation.
Conclusion
- The research verifies the initial hypothesis that the cryopreservation medium containing CM supports the cryopreservation of UCIM-MSCs and better maintains the biological characteristics observed in fresh cells.
- Further studies are encouraged to better understand the impact of the CM on UCIM-MSCs during freezing, shedding light on the beneficial and protective effects of using the CM during cryopreservation.
Cite This Article
APA
Maia L, Dias MC, de Moraes CN, de Paula Freitas-Dell'Aqua C, da Mota LS, Santiloni V, da Cruz Landim-Alvarenga F.
(2017).
Conditioned medium: a new alternative for cryopreservation of equine umbilical cord mesenchymal stem cells.
Cell Biol Int, 41(3), 239-248.
https://doi.org/10.1002/cbin.10708 Publication
Researcher Affiliations
- Department of Animal Reproduction, São Paulo State University, Prof. Doutor Walter Mauricio Correa n/n, CEP: 18618-681, Botucatu, São Paulo, Brazil.
- Department of Animal Reproduction, São Paulo State University, Prof. Doutor Walter Mauricio Correa n/n, CEP: 18618-681, Botucatu, São Paulo, Brazil.
- Department of Animal Reproduction, São Paulo State University, Prof. Doutor Walter Mauricio Correa n/n, CEP: 18618-681, Botucatu, São Paulo, Brazil.
- Department of Animal Reproduction, São Paulo State University, Prof. Doutor Walter Mauricio Correa n/n, CEP: 18618-681, Botucatu, São Paulo, Brazil.
- Department of Genetics, Bioscience Institute, São Paulo State University, Prof. Dr. Antônio Celso Wagner Zanin, 250, CEP: 18618-689, Botucatu, São Paulo, Brazil.
- Department of Genetics, Bioscience Institute, São Paulo State University, Prof. Dr. Antônio Celso Wagner Zanin, 250, CEP: 18618-689, Botucatu, São Paulo, Brazil.
- Department of Animal Reproduction, São Paulo State University, Prof. Doutor Walter Mauricio Correa n/n, CEP: 18618-681, Botucatu, São Paulo, Brazil.
MeSH Terms
- Animals
- Cell Differentiation / drug effects
- Cell Differentiation / physiology
- Cell Survival / drug effects
- Cell Survival / physiology
- Cells, Cultured
- Cryopreservation / methods
- Culture Media, Conditioned / pharmacology
- Horses
- Mesenchymal Stem Cells / drug effects
- Mesenchymal Stem Cells / physiology
- Umbilical Cord / cytology
- Umbilical Cord / drug effects
- Umbilical Cord / physiology
Citations
This article has been cited 1 times.- Nascimento C, Saraiva MVA, Pereira VM, de Brito DCC, de Aguiar FLN, Alves BG, Roballo KCS, de Figueiredo JR, Ambrósio CE, Rodrigues APR. Addition of synthetic polymer in the freezing solution of mesenchymal stem cells from equine adipose tissue as a future perspective for reducing of DMSO concentration. Braz J Vet Med 2023;45:e002523.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
