Translation of multipotent mesenchymal stromal cell (MSC)-based therapies is advancing
in human and veterinary medicine. One critical issue is the in vitro culture of MSC before
clinical use. Using fetal bovine serum (FBS) as supplement to the basal medium is still the
gold standard for cultivation of many cell types including equine MSC. Alternatives are
being explored, with substantial success using platelet lysate-supplemented media for
human MSC. However, progress lags behind in the veterinary field. The aim of this study
was to establish a scalable protocol for equine platelet lysate (ePL) production and to test
the ePL in equine MSC culture. Whole blood was harvested into blood collection bags
from 20 healthy horses. After checking sample materials for pathogen contamination,
samples from 19 animals were included. Platelet concentrates were prepared using a
buffy coat method. Platelets, platelet-derived growth factor BB, and transforming growth
factor b1 concentrations were increased in the concentrates compared with whole blood
or serum (p < 0.05), while white blood cells were reduced (p < 0.05). The concentrates
were lysed using freeze/thaw cycles, which eliminated the cells while growth factor
concentrations were maintained. Donor age negatively correlated with platelet and
growth factor concentrations after processing (p < 0.05). Finally, all lysates were pooled
and the ePL was evaluated as culture medium supplement in comparison with FBS,
using adipose-derived MSC from four unrelated donor horses. MSC proliferated well
in 10% FBS as well as in 10% ePL. However, using 5 or 2.5% ePL entailed highly
inconsistent proliferation or loss of proliferation, with significant differences in generation
times and confluencies (p < 0.05). MSC expressed the surface antigens CD90, CD44,
and CD29, but CD73 and CD105 detection was low in all culture media. Adipogenic
and osteogenic differentiation led to similar results in MSC from different culture media.
The buffy coat method is useful to produce equine platelet concentrate with increased
platelet and reduced white blood cell content in large scales. The ePL obtained supports
MSC expansion similar as FBS when used at the same concentration (10%). Further
investigations into equine MSC functionality in culture with ePL should follow.
| Identifer | oai:union.ndltd.org:DRESDEN/oai:qucosa:de:qucosa:84498 |
| Date | 03 April 2023 |
| Creators | Hagen, A., Lehmann, H., Aurich, S., Bauer, N., Melzer, M., Moellerberndt, J., Patané, V., Schnabel, C.L., Burk, J. |
| Publisher | Frontiers Research Foundation |
| Source Sets | Hochschulschriftenserver (HSSS) der SLUB Dresden |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, doc-type:article, info:eu-repo/semantics/article, doc-type:Text |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 2296-4185, 613621 |
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