Scalable Production of Equine Platelet Lysate for Multipotent Mesenchymal Stromal Cell Culture.
Scalable Production of Equine Platelet Lysate for Multipotent Mesenchymal Stromal Cell Culture.
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多能间充质基质细胞培养的马血小板裂解物的可扩展产生。
DOI：
10.3389/fbioe.2020.613621
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发表时间：
2020
影响因子：
5.7
通讯作者：
Burk J
中科院分区：
文献类型：
--
作者：
Hagen A;Lehmann H;Aurich S;Bauer N;Melzer M;Moellerberndt J;Patané V;Schnabel CL;Burk J
基于多能间充质基质细胞（MSC）的疗法的翻译在人类和兽医学中正在推进。一个关键问题是临床使用前MSC的体外培养。使用胎牛血清（FBS）作为基础培养基的补充仍然是培养包括马MSC在内的许多细胞类型的金标准。正在探索替代方案，使用补充血小板裂解物的人MSC培养基取得了实质性成功。然而，兽医领域的进展滞后。本研究的目的是建立一个可扩展的方案，马血小板裂解物（ePL）的生产和测试的ePL在马MSC培养。从20匹健康马采集全血至采血袋中。在检查样本材料的病原体污染后，纳入了19只动物的样本。使用血沉棕黄层方法制备血小板浓缩物。与全血或血清相比，浓缩物中的血小板、血小板衍生生长因子BB和转化生长因子β1浓度增加（p < 0.05），而白色血细胞减少（p < 0.05）。使用冻融循环裂解浓缩物，其消除细胞，同时保持生长因子浓度。供体年龄与处理后血小板和生长因子浓度呈负相关（p < 0.05）。最后，将所有裂解物合并，并使用来自四匹无关供体马的脂肪来源的MSC，将ePL作为培养基补充物与FBS进行比较。MSC在10%FBS和10%ePL中增殖良好。然而，使用5%或2.5%ePL导致高度不一致的增殖或增殖损失，在世代时间和汇合率方面具有显著差异（p < 0.05）。MSC表达表面抗原CD90，CD44和CD29，但CD73和CD105检测在所有培养基中均较低。成脂和成骨分化导致类似的结果在MSC从不同的培养基。血沉棕黄层法可用于大规模生产具有增加的血小板和减少的白色血细胞含量的马血小板浓缩物。当以相同浓度（10%）使用时，获得的ePL支持与FBS类似的MSC扩增。应进一步研究马MSC在ePL培养中的功能。
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 β1 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.
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影响因子：
7.5
作者：
Ben Azouna N;Jenhani F;Regaya Z;Berraeis L;Ben Othman T;Ducrocq E;Domenech J
通讯作者：
Domenech J
影响因子：
5.6
作者：
Doucet, C;Ernou, I;Lataillade, JJ
通讯作者：
Lataillade, JJ
影响因子：
2.9
作者：
Blande, Ivan Souza;Bassaneze, Vinicius;Krieger, Jose Eduardo
通讯作者：
Krieger, Jose Eduardo
影响因子：
4
作者：
Broeckx, Sarah Y.;Seys, Bert;Spaas, Jan H.
通讯作者：
Spaas, Jan H.
影响因子：
1
作者：
Braun, Julian;Hack, Anita;Aicher, Wilhelm K.
通讯作者：
Aicher, Wilhelm K.