Tissue Engineering Laboratories, BIOTEC and DFG Research Center and Cluster of Excellence for Regenerative Therapies Dresden (CRTD), Technische Universität Dresden, Dresden, Germany.
Cytotherapy. 2010 Apr;12(2):131-42. doi: 10.3109/14653240903476438.
It is unclear whether the plastic-adherent multipotent mesenchymal stromal cells (MSC) isolated from human bone marrow (BM) represent a uniform cell population or are heterogeneous in terms of cell-surface constituents and hence functionality.
We investigated the expression profile of certain biofunctional lipids by plastic-adherent MSC, focusing particularly on two membrane microdomain (lipid raft)-associated monosialogangliosides, GM1 and GM3, using indirect confocal laser scanning fluorescence microscopy and flow cytometry.
Phenotypically, we observed a differential expression where certain MSC subsets exhibited GM1, GM3 or both at the plasma membrane. Furthermore, disialoganglioside GD2 detection increased the complexity of the expression patterns, giving rise to seven identifiable cell phenotypes. Variation of standard culture conditions, such as the number of cell passage and period in culture, as well as donors, did not influence the heterologous ganglioside expression profile. In contrast, the binding of various lectins appeared homogeneous throughout the MSC population, indicating that the general glycosylation pattern remained common. Morphologically, the expression of a given ganglioside-based phenotype was not related to a cell with particular size or shape. Interestingly, a segregation of GM1 and GM3 clusters was observed, GM3 being mostly excluded from the highly curved plasma membrane protrusions.
These data highlight the phenotypic heterogeneity of plastic-adherent MSC in terms of certain lipid constituents of the plasma membrane, and the presence and/or absence of distinct ganglioside-based membrane microdomains suggest their potential functional diversity.
目前尚不清楚从人骨髓中分离出的黏附性多能间充质基质细胞(MSC)是否代表均匀的细胞群体,或者在细胞膜成分方面是否存在异质性,从而影响其功能。
我们使用间接共聚焦激光扫描荧光显微镜和流式细胞术,研究了黏附性 MSC 中某些生物功能脂质的表达谱,特别关注两种膜微区(脂质筏)相关的单唾液酸神经节苷脂 GM1 和 GM3。
在表型上,我们观察到某些 MSC 亚群在质膜上表现出 GM1、GM3 或两者均有差异表达。此外,双唾液酸神经节苷脂 GD2 的检测增加了表达模式的复杂性,产生了 7 种可识别的细胞表型。标准培养条件的变化,如细胞传代次数和培养时间以及供体,并不影响异源神经节苷脂表达谱。相比之下,各种凝集素的结合在 MSC 群体中表现出均匀性,表明一般的糖基化模式仍然是共同的。从形态上看,特定神经节苷脂表型的表达与具有特定大小或形状的细胞无关。有趣的是,观察到 GM1 和 GM3 簇的分离,GM3 主要不位于高度弯曲的质膜突起中。
这些数据突出了黏附性 MSC 在质膜某些脂质成分方面的表型异质性,以及特定神经节苷脂膜微区的存在和/或缺失表明其潜在的功能多样性。