Intracellular surface-enhanced Raman scattering probes based on TAT peptide-conjugated Au nanostars for distinguishing the differentiation of lung resident mesenchymal stem cells

基于 TAT 肽共轭金纳米星的细胞内表面增强拉曼散射探针用于区分肺驻留间充质干细胞的分化

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作者:Chaowen Shi, Xiaowei Cao, Xiang Chen, Zhaorui Sun, Zou Xiang, Hang Zhao, Weiping Qian, Xiaodong Han

Abstract

Lung resident mesenchymal stem cells (LR-MSCs) are important regulators of pathophysiological processes including tissue repair and fibrosis, inflammation, angiogenesis and tumor formation. Therefore, increasing attention has focused on the functional differentiation of LR-MSCs. However, the distinction between the undifferentiated and differentiated LR-MSCs, which are closely related and morphologically similar, is difficult to achieve by conventional methods. In this study, by employing the TAT Peptide-conjugated Au nanostars (AuNSs) as an intracellular probe, we developed a method for the identification of LR-MSC differentiation by surface-enhanced Raman scattering (SERS) spectroscopy. SERS spectra were analyzed using principal component analysis (PCA) that allowed unambiguous distinction of subtypes and monitoring of component changes during cellular differentiation. Furthermore, to ascertain whether co-culture with alveolar epithelial type II (ATII) cells and incubation with transform growth factor (TGF)-β were involved in regulating the differentiation of LR-MSCs, we investigated the protein expression levels of epithelial markers and fibroblastic markers on LR-MSCs. Our results demonstrated that co-culture with ATII cells or incubation with TGF-β could induce the differentiation of LR-MSCs as confirmed by SERS analysis, a method that is capable of noninvasive characterization of and distinction between subtypes of LR-MSCs during differentiation. We have provided a new tool that may facilitate stem cell research.

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