Canine and Equine Mesenchymal Stem Cells Grown in Serum Free Media Have Altered Immunophenotype
Canine and Equine Mesenchymal Stem Cells Grown in Serum Free Media Have Altered Immunophenotype
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DOI：
10.1007/s12015-015-9638-0
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发表时间：
2016-04-01
影响因子：
4.8
通讯作者：
Borjesson, Dori L.
中科院分区：
文献类型：
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作者：
Clark, Kaitlin C.;Kol, Amir;Borjesson, Dori L.
间充质干细胞（MSC）疗法越来越多地用于治疗患有自然发生疾病的狗和马。然而，这些动物也作为关键的大型动物模型，用于将细胞治疗产品持续转化为人类市场。用于临床用途的MSC制造要求改进细胞培养系统以满足对更高MSC数量和去除异种蛋白（即胎牛血清，FBS）的需求。虽然无血清培养基（SFM）是可商购的，但其对MSC表型和免疫调节功能的影响尚不完全清楚。本研究的目的是确定特定的MSC培养条件，即在HYPERFlasksA中的MSC扩增或在市售SFM中的MSC扩增，是否会改变MSC的体外增殖、表型或免疫调节特性。在HYPERFlasksA（R）中培养的MSC在表型、增殖能力和免疫调节功能方面与在标准烧瓶中生长的MSC相似，但是MSC产量显著增加。因此，HYPERFlasksA（R）提供了一个可行的选择，以产生更大的细胞数量在一个流线型的方式。犬和马骨髓间充质干细胞在SFM中体外扩增后，其增殖、表面表型和对淋巴细胞增殖的抑制作用相似。然而，在不存在FBS的情况下培养的MSC分泌显著较少的PGE（2），并且显著较少能够抑制活化的T细胞分泌IFN γ。免疫调节功能改变的扩展SFM种属依赖性。与马MSC不同，在犬脂肪来源的MSC中，淋巴细胞增殖的抑制主要不受PGE调节（2）。从犬和马MSC培养系统中去除FBS导致体外免疫调节特性改变，并且在转向无FBS培养条件之前需要进一步研究。
Mesenchymal stem cell (MSC) therapy is being increasingly used to treat dogs and horses with naturally-occurring diseases. However these animals also serve as critical large animal models for ongoing translation of cell therapy products to the human market. MSC manufacture for clinical use mandates improvement in cell culture systems to meet demands for higher MSC numbers and removal of xeno-proteins (i.e. fetal bovine serum, FBS). While serum-free media (SFM) is commercially available, its affects on MSC phenotype and immunomodulatory functions are not fully known. The objective of this study was to determine if specific MSC culture conditions, MSC expansion in HYPERFlasksA (R) or MSC expansion in a commercially available SFM, would alter MSC proliferation, phenotype or immunomodulatory properties in vitro. MSCs cultured in HYPERFlasksA (R) were similar in phenotype, proliferative capacity and immunomodulatory functions to MSCs grown in standard flasks however MSC yield was markedly increased. HYPERFlasksA (R) therefore provide a viable option to generate greater cell numbers in a streamlined manner. Canine and equine MSCs expanded in SFM displayed similar proliferation, surface phenotype and inhibitory effect on lymphocyte proliferation in vitro. However, MSCs cultured in the absence of FBS secreted significantly less PGE(2), and were significantly less able to inhibit IFN gamma secretion by activated T-cells. Immunomodulatory functions altered by expansion in SFM were species dependent. Unlike equine MSCs, in canine adipose-derived MSCs, the inhibition of lymphocyte proliferation was not principally modulated by PGE(2). The removal of FBS from both canine and equine MSC culture systems resulted in altered immunomodulatory properties in vitro and warrants further investigation prior to moving towards FBS-free culture conditions.