Serum‐free human MSC medium supports consistency in human but not in equine adipose‐derived multipotent mesenchymal stromal cell culture
Serum‐free human MSC medium supports consistency in human but not in equine adipose‐derived multipotent mesenchymal stromal cell culture
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无血清人 MSC 培养基支持人类但不支持马脂肪源多能间充质基质细胞培养的一致性
DOI：
10.1002/cyto.a.23240
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Burk J.
中科院分区：
文献类型：
--
作者：
Schubert S;Brehm W;Hillmann A;Burk J.
对于多能间充质基质细胞（MSC）的临床应用，无血清培养是优选的，以标准化细胞产品并防止病原体污染。与人MSC相反，关于大型动物MSC的无血清培养的知识是有限的，尽管其与临床前研究和兽医细胞疗法的开发相关。本研究旨在评价市售无血清人MSC培养基与人脂肪MSC相比培养马脂肪来源MSC的适用性。通过外植体技术进行无酶分离以及在无血清培养基中扩增马和人细胞是可行的。然而，与在补充有胎牛血清的标准培养基中培养相比，无血清培养改变了马MSC的形态和复杂的处理，具有细胞聚集和多层自发脱离。此外，与对照组相比，马细胞的增殖和表面免疫表型变化更大，似乎取决于无血清培养基的批次。特别是实验组之间CD 90的表达不同（P< 0.05），与标准培养基（86.20-99.50%）相比，在无血清培养基（5.21-83.40%）中培养的马MSC样品中发现的CD 90+细胞百分比较低。此外，在无血清培养后的马样本中发现了表达MSC排除标志物的小亚群，如CD 14（0.28-11.60%）、CD 34（0.00-9.87%）、CD 45（0.35-10.50%）或MHCII（0.00-3.67%）。相比之下，在无血清培养基中培养后，人样本显示出更一致的形态学和一致的CD 29+（98.60-99.90%）、CD 73+（94.60 -98.40%）、CD 90+（99.60-99.90%）和CD 105+（97.40-99.80%）免疫表型。获得的数据表明，无血清培养基适用于人MSC培养，但在马MSC中并未产生完全令人满意的结果。这强调了关于无血清培养条件的要求是物种特异性的，表明需要为来自相关动物物种的MSC优化无血清培养基。© 2017国际细胞计数促进学会
For clinical applications of multipotent mesenchymal stromal cells (MSCs), serum‐free culture is preferable to standardize cell products and prevent contamination with pathogens. In contrast to human MSCs, knowledge on serum‐free culture of large animal MSCs is limited, despite its relevance for preclinical studies and development of veterinary cellular therapeutics. This study aimed to evaluate the suitability of a commercially available serum‐free human MSC medium for culturing equine adipose‐derived MSCs in comparison with human adipose MSCs. Enzyme‐free isolation by explant technique and expansion of equine and human cells in the serum‐free medium were feasible. However, serum‐free culture altered the morphology and complicated handling of equine MSCs, with cell aggregation and spontaneous detachment of multilayers, compared to culture in standard medium supplemented with fetal bovine serum. Furthermore, proliferation and the surface immunophenotype of equine cells were more variable compared to the controls and appeared to depend on the lot of the serum‐free medium. Particularly the expression of CD90 was different between experimental groups (P< 0.05), with lower percentages of CD90+cells found in equine MSC samples cultured in serum‐free medium (5.21–83.40%) compared to standard medium (86.20–99.50%). Additionally, small subpopulations expressing MSC exclusion markers such as CD14 (0.28–11.60%), CD34 (0.00–9.87%), CD45 (0.35–10.50%), or MHCII (0.00–3.67%) were found in equine samples after serum‐free culture. In contrast, human samples displayed a more consistent morphology and a consistent CD29+(98.60–99.90%), CD73+(94.60–98.40%), CD90+(99.60–99.90%), and CD105+(97.40–99.80%) immunophenotype after culture in serum‐free medium. The obtained data demonstrate that the serum‐free medium was suitable for human MSC culture but did not lead to entirely satisfactory results in equine MSCs. This underlines that requirements regarding serum‐free culture conditions are species‐specific, indicating a need for serum‐free media to be optimized for MSCs from relevant animal species. © 2017 International Society for Advancement of Cytometry
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通讯作者：
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影响因子：
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