Phospholipid Profiles for Phenotypic Characterization of Adipose-Derived Multipotent Mesenchymal Stromal Cells.
Phospholipid Profiles for Phenotypic Characterization of Adipose-Derived Multipotent Mesenchymal Stromal Cells.
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DOI：
10.3389/fcell.2021.784405
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发表时间：
2021
影响因子：
5.5
通讯作者：
Leopold J
中科院分区：
文献类型：
--
作者：
Burk J;Melzer M;Hagen A;Lips KS;Trinkaus K;Nimptsch A;Leopold J
多能间充质基质细胞（MSC）已成为广泛的病理条件的治疗工具。然而，关于MSC表型表征的仍然存在的缺陷以及在不同的临床前和临床研究中使用的MSC的异质性阻碍了转化的成功。在寻找新的MSC表征方法，以补充传统的三系分化和免疫表型分析可靠的跨物种和培养条件下，本研究探讨了脂质表型MSC表征和歧视的适用性。人外周血单核细胞（PBMC），人成纤维细胞，人和马脂肪来源的MSC用于比较不同的中胚层细胞类型和不同物种的MSC。对于MSC，另外研究了在不同条件下培养的细胞，包括补充有胎牛血清或血小板裂解物的培养基以及在胶原包被的培养皿上培养。细胞收获后，根据Bligh和Dyer通过氯仿/甲醇提取脂质。通过非靶向方法，使用液相色谱-质谱联用（LC-MS）（配有反相柱和离子阱质谱仪）分析脂质谱。在所有样品中，发现磷脂和鞘磷脂，而其他脂质没有检测到与当前的方法。在所有细胞类型中，磷脂包括不同种类的磷脂酰胆碱（PC）、磷脂酰乙醇胺（PE）、磷脂酰肌醇（PI）和磷脂酰丝氨酸（PS），而磷脂酰甘油（PG）种类仅存在于MSC中。来自两个物种的MSC显示出比PBMC和成纤维细胞更高的磷脂种类多样性。在来自不同培养条件的MSC之间发现很少的差异，除了用血小板裂解物培养的人MSC表现出独特的表型，因为它们仅以PE 0 -40：4、PG 38：6和PG 40：6为特征。在寻找特异性和包容性的候选MSC脂质标志物时，我们将PE O-36：3和PG 40：7鉴定为跨培养条件的潜在合适标志物，其中PE O-36：3甚至可以跨物种使用。在此基础上，磷脂表型是一种非常有前途的MSC表征方法，它可能会容忍MSC内的一些异质性，同时仍然实现了甚至与成纤维细胞的明确区分。特别是PG的存在或不存在可能成为未来MSC表征的决定性标准。
Multipotent mesenchymal stromal cells (MSC) have emerged as therapeutic tools for a wide range of pathological conditions. Yet, the still existing deficits regarding MSC phenotype characterization and the resulting heterogeneity of MSC used in different preclinical and clinical studies hamper the translational success. In search for novel MSC characterization approaches to complement the traditional trilineage differentiation and immunophenotyping assays reliably across species and culture conditions, this study explored the applicability of lipid phenotyping for MSC characterization and discrimination. Human peripheral blood mononuclear cells (PBMC), human fibroblasts, and human and equine adipose-derived MSC were used to compare different mesodermal cell types and MSC from different species. For MSC, cells cultured in different conditions, including medium supplementation with either fetal bovine serum or platelet lysate as well as culture on collagen-coated dishes, were additionally investigated. After cell harvest, lipids were extracted by chloroform/methanol according to Bligh and Dyer. The lipid profiles were analysed by an untargeted approach using liquid chromatography coupled to mass spectrometry (LC-MS) with a reversed phase column and an ion trap mass spectrometer. In all samples, phospholipids and sphingomyelins were found, while other lipids were not detected with the current approach. The phospholipids included different species of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and phosphatidylserine (PS) in all cell types, whereas phosphatidylglycerol (PG) species were only present in MSC. MSC from both species showed a higher phospholipid species diversity than PBMC and fibroblasts. Few differences were found between MSC from different culture conditions, except that human MSC cultured with platelet lysate exhibited a unique phenotype in that they exclusively featured PE O-40:4, PG 38:6 and PG 40:6. In search for specific and inclusive candidate MSC lipid markers, we identified PE O-36:3 and PG 40:7 as potentially suitable markers across culture conditions, at which PE O-36:3 might even be used across species. On that basis, phospholipid phenotyping is a highly promising approach for MSC characterization, which might condone some heterogeneity within the MSC while still achieving a clear discrimination even from fibroblasts. Particularly the presence or absence of PG might emerge as a decisive criterion for future MSC characterization.
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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。
影响因子：
5.7
作者：
Hagen A;Lehmann H;Aurich S;Bauer N;Melzer M;Moellerberndt J;Patané V;Schnabel CL;Burk J
通讯作者：
Burk J
影响因子：
5.4
作者：
Klein ME;Rieckmann M;Sedding D;Hause G;Meister A;Mäder K;Lucas H
通讯作者：
Lucas H
影响因子：
6.5
作者：
Kilpinen, Lotta;Tigistu-Sahle, Feven;Laitinen, Saara
通讯作者：
Laitinen, Saara
影响因子：
1.9
作者：
Engel, Kathrin M.;Schiller, Juergen;Jakop, Ulrike
通讯作者：
Jakop, Ulrike
影响因子：
3
作者：
Frith, Jessica E.;Thomson, Brian;Genever, Paul G.
通讯作者：
Genever, Paul G.