Carlomagno Cristiano, Giannasi Chiara, Niada Stefania, Bedoni Marzia, Gualerzi Alice, Brini Anna Teresa
IRCCS Fondazione Don Carlo Gnocchi ONLUS, Milan, Italy.
Department of Biomedical Surgical and Dental Sciences, University of Milan, Milan, Italy.
Front Bioeng Biotechnol. 2021 Apr 13;9:640617. doi: 10.3389/fbioe.2021.640617. eCollection 2021.
Extracellular Vesicles (EVs) and Conditioned Medium (CM) are promising cell-free approaches to repair damaged and diseased tissues for regenerative rehabilitation purposes. They both entail several advantages, mostly in terms of safety and handling, compared to the cell-based treatment. Despite the growing interest in both EVs and CM preparations, in the light of a clinical translation, a number of aspects still need to be addressed mainly because of limits in the reproducibility and reliability of the proposed protocols. Raman spectroscopy (RS) is a non-destructive vibrational investigation method that provides detailed information about the biochemical composition of a sample, with reported ability in bulk characterization of clusters of EVs from different cell types. In the present brief report, we acquired and compared the Raman spectra of the two most promising cell-free therapeutics, i.e., EVs and CM, derived from two cytotypes with a history in the field of regenerative medicine, adipose-derived mesenchymal stem/stromal cells (ASCs) and dermal fibroblasts (DFs). Our results show how RS can verify the reproducibility not only of EV isolation, but also of the whole CM, thus accounting for both the soluble and the vesicular components of cell secretion. RS can provide hints for the identification of the soluble factors that synergistically cooperate with EVs in the regenerative effect of CM. Still, we believe that the application of RS in the pipeline of cell-free products preparation for therapeutic purposes could help in accelerating translation to clinics and regulatory approval.
细胞外囊泡（EVs）和条件培养基（CM）是很有前景的无细胞方法，用于修复受损和患病组织以实现再生康复目的。与基于细胞的治疗相比，它们都有几个优点，主要体现在安全性和操作方面。尽管对EVs和CM制剂的兴趣日益增加，但鉴于临床转化，仍有许多方面需要解决，主要是因为所提出方案的可重复性和可靠性存在局限性。拉曼光谱（RS）是一种非破坏性振动研究方法，可提供有关样品生化组成的详细信息，据报道能够对来自不同细胞类型的EVs簇进行整体表征。在本简要报告中，我们获取并比较了两种最有前景的无细胞疗法（即EVs和CM）的拉曼光谱，这两种疗法分别源自再生医学领域有相关历史的两种细胞类型，即脂肪来源的间充质干/基质细胞（ASCs）和真皮成纤维细胞（DFs）。我们的结果表明，RS不仅可以验证EV分离的可重复性，还可以验证整个CM的可重复性，从而兼顾了细胞分泌的可溶性成分和囊泡成分。RS可以为识别在CM的再生作用中与EVs协同作用的可溶性因子提供线索。尽管如此，我们认为RS在用于治疗目的的无细胞产品制备流程中的应用有助于加速向临床转化和监管审批。
