Regenerative Medicine Group, Department of Health Science and Technology, Aalborg University, Fredrik Bajers Vej 3B, 9220 Aalborg, Denmark.
Cells. 2021 Jan 22;10(2):218. doi: 10.3390/cells10020218.
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 (CD166CD34CD146CD271 CD274CD248CD200 and CD166CD34 CD146CD271CD274CD248CD200) and one clone in the smaller panel (CD29CD201CD36 Stro-1 CD31). The minor subsets, including CD166CD34CD146CD271CD274CD248CD200 and CD166CD34CD146CD271CD274CD248CD200, and CD29CD201CD36Stro-1CD31, CD29CD201CD36Stro-1CD31, and CD29CD201CD36Stro-1CD31, 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.
为了提高治疗效果，了解脂肪干细胞（ASC）在体外生长过程中免疫表型和生物学特性的动态变化非常重要。因此，我们着手研究从三个无关供体在组织培养聚苯乙烯上进行的八次传代过程中，高度定义的细胞亚群的演变。通过包含七个和五个细胞表面标志物的面板定义共表达模式，分别包括 CD34、CD146、CD166、CD200、CD248、CD271 和 CD274 和 CD29、CD31、CD36、CD201 和 Stro-1。分析使用多色流式细胞术进行。我们观察到一个主要的范式转变，其中在培养早期在所有细胞亚群中发现的 CD166-CD34 组合被 CD166 表型取代，因为随着时间的推移，群体均一性增加。在所有分析点，培养物都由少数几个主要克隆主导，这些克隆在大多数供体中都非常普遍。选择过程导致在较大面板（CD166CD34CD146CD271CD274CD248CD200 和 CD166CD34 CD146CD271CD274CD248CD200）中出现两个主要克隆，而在较小面板（CD29CD201CD36Stro-1CD31）中出现一个克隆。包括 CD166CD34CD146CD271CD274CD248CD200 和 CD166CD34CD146CD271CD274CD248CD200 在内的较小亚群，以及 CD29CD201CD36Stro-1CD31、CD29CD201CD36Stro-1CD31 和 CD29CD201CD36Stro-1CD31，在七个和五个标记面板中，分别为，另一方面，高度波动且供体依赖性。结果表明，ASC 培养物中可能只有有限数量的表型谱。不同个体之间相对发生的显著差异强调了需要对不同 ASC 制剂进行效力标准化，以提高临床疗效。
