Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells
Abstract Background Mesenchymal stem cells (MSCs) produced for clinical purposes rely on culture media containing fetal bovine serum (FBS) which is xenogeneic and has the potential to significantly alter the MSC phenotype, rendering these cells immunogenic. As a result of bovine-derived exogenous pr...
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| Format: | Article |
| Language: | English |
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BMC
2018-03-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13287-018-0823-3 |
| _version_ | 1818469136734355456 |
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| author | Maria C. Naskou Scarlett M. Sumner Anna Chocallo Hannah Kemelmakher Merrilee Thoresen Ian Copland Jacques Galipeau John F. Peroni |
| author_facet | Maria C. Naskou Scarlett M. Sumner Anna Chocallo Hannah Kemelmakher Merrilee Thoresen Ian Copland Jacques Galipeau John F. Peroni |
| author_sort | Maria C. Naskou |
| collection | DOAJ |
| description | Abstract Background Mesenchymal stem cells (MSCs) produced for clinical purposes rely on culture media containing fetal bovine serum (FBS) which is xenogeneic and has the potential to significantly alter the MSC phenotype, rendering these cells immunogenic. As a result of bovine-derived exogenous proteins expressed on the cell surface, MSCs may be recognized by the host immune system as non-self and be rejected. Platelet lysate (PL) may obviate some of these concerns and shows promising results in human medicine as a possible alternative to FBS. Our goal was to evaluate the use of equine platelet lysate (ePL) pooled from donor horses in place of FBS to culture equine MSCs. We hypothesized that ePL, produced following apheresis, will function as the sole media supplement to accelerate the expansion of equine bone marrow-derived MSCs without altering their phenotype and their immunomodulatory capacity. Methods Platelet concentrate was obtained via plateletpheresis and ePL were produced via freeze-thaw and centrifugation cycles. Population doublings (PD) and doubling time (DT) of bone marrow-derived MSCs (n = 3) cultured with FBS or ePL media were calculated. Cell viability, immunophenotypic analysis, and trilineage differentiation capacity of MSCs were assessed accordingly. To assess the ability of MSCs to modulate inflammatory responses, E. coli lipopolysaccharide (LPS)-stimulated monocytes were cocultured with MSCs cultured in the two different media formulations, and cell culture supernatants were assayed for the production of tumor necrosis factor (TNF)-α. Results Our results showed that MSCs cultured in ePL media exhibited similar proliferation rates (PD and DT) compared with those cultured in FBS at individual time points. MSCs cultured in ePL showed a statistically significant increased viability following a single washing step, expressed similar levels of MSC markers compared to FBS, and were able to differentiate towards the three lineages. Finally, MSCs cultured in ePL efficiently suppressed the release of TNF-α when exposed to LPS-stimulated monocytes similar to those cultured in FBS. Conclusion ePL has the potential to be used for the expansion of MSCs before clinical application, avoiding the concerns associated with the use of FBS. |
| first_indexed | 2024-04-13T21:20:59Z |
| format | Article |
| id | doaj.art-fcd5f8f9b255476282d9b9951053f2d2 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-04-13T21:20:59Z |
| publishDate | 2018-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-fcd5f8f9b255476282d9b9951053f2d22022-12-22T02:29:31ZengBMCStem Cell Research & Therapy1757-65122018-03-019111310.1186/s13287-018-0823-3Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cellsMaria C. Naskou0Scarlett M. Sumner1Anna Chocallo2Hannah Kemelmakher3Merrilee Thoresen4Ian Copland5Jacques Galipeau6John F. Peroni7Department of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of GeorgiaDepartment of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of GeorgiaDepartment of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of GeorgiaDepartment of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of GeorgiaDepartment of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of GeorgiaEmory Personalized Immunotherapy Center [EPIC], Emory University School of MedicineDepartment of Medicine and Carbone Comprehensive Cancer Center, University of WisconsinDepartment of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of GeorgiaAbstract Background Mesenchymal stem cells (MSCs) produced for clinical purposes rely on culture media containing fetal bovine serum (FBS) which is xenogeneic and has the potential to significantly alter the MSC phenotype, rendering these cells immunogenic. As a result of bovine-derived exogenous proteins expressed on the cell surface, MSCs may be recognized by the host immune system as non-self and be rejected. Platelet lysate (PL) may obviate some of these concerns and shows promising results in human medicine as a possible alternative to FBS. Our goal was to evaluate the use of equine platelet lysate (ePL) pooled from donor horses in place of FBS to culture equine MSCs. We hypothesized that ePL, produced following apheresis, will function as the sole media supplement to accelerate the expansion of equine bone marrow-derived MSCs without altering their phenotype and their immunomodulatory capacity. Methods Platelet concentrate was obtained via plateletpheresis and ePL were produced via freeze-thaw and centrifugation cycles. Population doublings (PD) and doubling time (DT) of bone marrow-derived MSCs (n = 3) cultured with FBS or ePL media were calculated. Cell viability, immunophenotypic analysis, and trilineage differentiation capacity of MSCs were assessed accordingly. To assess the ability of MSCs to modulate inflammatory responses, E. coli lipopolysaccharide (LPS)-stimulated monocytes were cocultured with MSCs cultured in the two different media formulations, and cell culture supernatants were assayed for the production of tumor necrosis factor (TNF)-α. Results Our results showed that MSCs cultured in ePL media exhibited similar proliferation rates (PD and DT) compared with those cultured in FBS at individual time points. MSCs cultured in ePL showed a statistically significant increased viability following a single washing step, expressed similar levels of MSC markers compared to FBS, and were able to differentiate towards the three lineages. Finally, MSCs cultured in ePL efficiently suppressed the release of TNF-α when exposed to LPS-stimulated monocytes similar to those cultured in FBS. Conclusion ePL has the potential to be used for the expansion of MSCs before clinical application, avoiding the concerns associated with the use of FBS.http://link.springer.com/article/10.1186/s13287-018-0823-3Equine platelet apheresisEquine platelet lysateMesenchymal stem cellsFetal bovine serumCell culture |
| spellingShingle | Maria C. Naskou Scarlett M. Sumner Anna Chocallo Hannah Kemelmakher Merrilee Thoresen Ian Copland Jacques Galipeau John F. Peroni Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells Stem Cell Research & Therapy Equine platelet apheresis Equine platelet lysate Mesenchymal stem cells Fetal bovine serum Cell culture |
| title | Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells |
| title_full | Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells |
| title_fullStr | Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells |
| title_full_unstemmed | Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells |
| title_short | Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells |
| title_sort | platelet lysate as a novel serum free media supplement for the culture of equine bone marrow derived mesenchymal stem cells |
| topic | Equine platelet apheresis Equine platelet lysate Mesenchymal stem cells Fetal bovine serum Cell culture |
| url | http://link.springer.com/article/10.1186/s13287-018-0823-3 |
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