Koczkowska Magdalena, Kostecka Anna, Zawrzykraj Małgorzata, Myszczyński Kamil, Skoniecka Aneta, Deptuła Milena, Tymińska Agata, Czerwiec Katarzyna, Jąkalski Marcin, Zieliński Jacek, Crossman David K, Crowley Michael R, Cichorek Mirosława, Skowron Piotr M, Pikuła Michał, Piotrowski Arkadiusz
3P-Medicine Laboratory, Medical University of Gdansk, Gdansk, Poland.
Department of Biology and Pharmaceutical Botany, Medical University of Gdansk, Gdansk, Poland.
Stem Cell Res Ther. 2025 Feb 11;16(1):64. doi: 10.1186/s13287-025-04185-w.
Adipose-derived mesenchymal stromal cells (AD-MSCs) and fibroblasts are both widely used in regenerative medicine, demonstrating significant potential for personalized cell therapy. A major challenge in their use lies in their high biological similarity, encompassing morphology, differentiation capabilities, and flow cytometric markers, making their distinction difficult.
In our study, we aimed to compare AD-MSCs obtained from two types of adipose tissue, subcutaneous and visceral, alongside skin fibroblasts. Notably, all tissue samples were sourced from the same donors. We analyzed the cells for surface antigens via flow cytometry and conducted single-cell RNA sequencing, followed by verification with quantitative PCR (qPCR).
Our results revealed phenotypic similarities between the isolated AD-MSCs and dermal fibroblasts, particularly in the expression of markers characteristic of AD-MSCs. However, through in-depth analyses, we identified distinct differences between these cell types. Specifically, we pinpointed 30 genes exhibiting the most significant variations in expression between AD-MSCs and fibroblasts. These genes are associated with biological processes such as tissue remodeling, cell movement, and activation in response to external stimuli. Among them, MMP1, MMP3, S100A4, CXCL1, PI16, IGFBP5, COMP were further validated using qPCR, clearly demonstrating their potential to differentiate between AD-MSCs and fibroblasts.
Our scRNA-seq analysis elucidates the transcriptional landscape of AD-MSCs and fibroblasts with unprecedented resolution, highlighting both the population-specific markers and the intrapopulation heterogeneity. Our findings underscore the importance of employing high-resolution techniques for cell identification.
脂肪来源的间充质基质细胞（AD-MSCs）和成纤维细胞都广泛应用于再生医学，显示出个性化细胞治疗的巨大潜力。它们使用中的一个主要挑战在于其高度的生物学相似性，包括形态、分化能力和流式细胞术标记，这使得它们难以区分。
在我们的研究中，我们旨在比较从皮下和内脏两种脂肪组织中获得的AD-MSCs以及皮肤成纤维细胞。值得注意的是，所有组织样本均来自相同的供体。我们通过流式细胞术分析细胞的表面抗原，并进行单细胞RNA测序，随后用定量PCR（qPCR）进行验证。
我们的结果揭示了分离出的AD-MSCs与真皮成纤维细胞之间的表型相似性，特别是在AD-MSCs特征性标记的表达方面。然而，通过深入分析，我们确定了这些细胞类型之间的明显差异。具体而言，我们确定了30个在AD-MSCs和成纤维细胞之间表达差异最为显著的基因。这些基因与组织重塑、细胞运动和对外界刺激的激活等生物学过程相关。其中，MMP1、MMP3、S100A4、CXCL1、PI16、IGFBP5、COMP通过qPCR进一步验证，清楚地表明它们在区分AD-MSCs和成纤维细胞方面的潜力。
我们的scRNA-seq分析以前所未有的分辨率阐明了AD-MSCs和成纤维细胞的转录图谱，突出了群体特异性标记和群体内异质性。我们的发现强调了采用高分辨率技术进行细胞鉴定的重要性。
