In order to enhance the therapeutic potential, it is important that sufficient knowledge regarding the dynamic changes of adipose-derived stem cell (ASC) immunophenotypical and biological properties during in vitro growth is available. Consequently, we embarked on a study to follow the evolution of highly defined cell subsets from three unrelated donors in the course of eight passages on tissue culture polystyrene. The co-expression patterns were defined by panels encompassing seven and five cell surface markers, including CD34, CD146, CD166, CD200, CD248, CD271, and CD274 and CD29, CD31, CD36, CD201, and Stro-1, respectively. The analysis was performed using multichromatic flow cytometry. We observed a major paradigm shift, where the CD166-CD34⁺ combination which was found across all cell subsets early in the culture was replaced by the CD166⁺ phenotype as the population homogeneity increased with time. At all analysis points, the cultures were dominated by a few major clones that were highly prevalent in most of the donors. The selection process resulted in two predominant clones in the larger panel (CD166⁺CD34⁻CD146⁻CD271⁻ CD274⁻CD248⁻CD200⁻ and CD166⁺CD34⁺ CD146⁻CD271⁻CD274⁻CD248⁻CD200⁻) and one clone in the smaller panel (CD29⁺CD201⁺CD36⁻ Stro-1⁻ CD31⁻). The minor subsets, including CD166⁺CD34⁻CD146⁻CD271⁺CD274⁻CD248⁻CD200⁻ and CD166⁺CD34⁺CD146⁺CD271⁻CD274⁻CD248⁻CD200⁻, and CD29⁺CD201⁻CD36⁻Stro-1⁻CD31⁻, CD29⁺CD201⁺CD36⁻Stro-1⁺CD31⁻, and CD29⁺CD201⁺CD36⁺Stro-1⁻CD31⁻, in the seven and five marker panels, respectively, were, on the other, hand highly fluctuating and donor-dependent. The results demonstrate that only a limited number of phenotypical repertoires are possible in ASC cultures. Marked differences in their relative occurrence between distinct individuals underscore the need for potency standardization of different ASC preparation to improve the clinical outcome.

中文翻译：

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脂肪衍生干细胞 (ASC) 免疫表型适应体外扩增的多重分析

为了提高治疗潜力，重要的是要有足够的关于体外生长过程中脂肪源性干细胞 (ASC) 免疫表型和生物学特性的动态变化的知识。因此，我们开始了一项研究，以追踪在组织培养聚苯乙烯的八次传代过程中来自三个无关供体的高度确定的细胞亚群的进化。共表达模式由包含七个和五个细胞表面标记的面板定义，分别包括 CD34、CD146、CD166、CD200、CD248、CD271 和 CD274 以及 CD29、CD31、CD36、CD201 和 Stro-1。使用多色流式细胞术进行分析。我们观察到一个主要的范式转变，其中 CD166-CD34 ⁺随着群体同质性随时间增加，在培养早期在所有细胞亚群中发现的组合被 CD166 ⁺表型取代。在所有分析点，培养物以少数主要克隆为主，这些克隆在大多数供体中高度流行。选择过程导致较大面板中的两个主要克隆（CD166 ⁺ CD34 ⁻ CD146 ⁻ CD271 ⁻ CD274 ⁻ CD248 ⁻ CD200 ⁻和 CD166 ⁺ CD34 ⁺ CD146 ⁻ CD271 ⁻ CD274 ⁻ CD248 ⁻ CD200 ⁻) 和较小面板中的一个克隆 (CD29 ⁺ CD201 ⁺ CD36 ⁻ Stro-1 ⁻ CD31 ⁻ )。次要子集，包括CD166 ⁺ CD34 ^- CD146 ^- CD271 ⁺ CD274 ^- CD248 ^- CD200 ^-和CD166 ⁺ CD34 ⁺ CD146 ⁺ CD271 ^- CD274 ^- CD248 ^- CD200 ^-，和CD29 ⁺ CD201 ^- CD36 ^-的Stro-1 ^- CD31 ^-、CD29 ⁺ CD201 ⁺ CD36 ^- Stro-1 ⁺ CD31 ^-和 CD29 ⁺ CD201 ⁺ CD36 ⁺ Stro-1 ^- CD31 ^{- 分别}在七个和五个标记组中，另一方面，高度波动和供体-依赖。结果表明，在 ASC 培养物中可能只有有限数量的表型曲目。不同个体之间相对发生率的显着差异强调了对不同 ASC 制剂进行效力标准化以改善临床结果的必要性。