在组织工程中，实时监测肿瘤和体外模型中微环境的动态传统上一直受到收获培养物以获得分析材料的需要的阻碍。线场共焦光学相干断层扫描 (LC-OCT) 已被证明可通过捕获动态三维 (3D) 信息来评估体内皮肤状况（包括黑色素瘤），而无需进行活检等侵入性操作。此外，IMcoMET 开发的 M-Duo Technology® 为连续原位生物标志物采样提供了独特的机会，提供了对局部细胞行为和相互作用的见解。本研究旨在验证两种先进方法（LC-OCT由 DAMAE Medical 和 M-Duo Technology® by IMcoMET）研究体外重建人类皮肤 (RhS) 和黑色素瘤-RhS (Mel-RhS) 的生活微环境。LC-OCT 和 M-Duo Technology® 与常规分析进行比较RhS 和 Mel-RhS 微环境的分析。LC-OCT 成功地可视化了 Mel-RhS 内表皮和肿瘤结构的不同层，识别了角质形成细胞、黑素细胞、肿瘤巢和成纤维细胞。 M-Duo Technology® 揭示了 Mel-RhS 和对照 RhS 之间原位细胞因子 (IL-6) 和趋化因子（CCL2、CXCL10 和 IL-8）分泌的差异。值得注意的是，通过对培养物上清液中分泌蛋白的常规研究无法检测到这种差异。LC-OCT 的高分辨率成像和 M-Duo Technology® 的原位微环境图谱的结合有可能为非侵入式实时分析，允许长时间观察 Mel-RhS 模型中的过程，无需进行培养破坏。版权所有 © 2024 作者。由 Elsevier B.V. 出版。保留所有权利。

In tissue engineering, real-time monitoring of tumors and of the dynamics of the microenvironment within in vitro models has traditionally been hindered by the need to harvest the cultures to obtain material to analyze. Line-field confocal optical coherence tomography (LC-OCT) has proven to be useful in evaluating in vivo skin conditions, including melanoma, by capturing dynamic, three-dimensional (3D) information without the need for invasive procedures, such as biopsies. Additionally, the M-Duo Technology® developed by IMcoMET presents a unique opportunity for continuous in situ biomarker sampling, providing insights into local cellular behavior and interactions.This study aimed to validate the non-destructive mapping capabilities of two advanced methodologies (LC-OCT by DAMAE Medical and M-Duo Technology® by IMcoMET) to investigate the living microenvironment of in vitro reconstructed human skin (RhS) and melanoma-RhS (Mel-RhS).LC-OCT and M-Duo Technology® were compared to conventional analysis of the RhS and Mel-RhS microenvironments.LC-OCT successfully visualized the distinct layers of the epidermis and tumor structures within the Mel-RhS, identifying keratinocytes, melanocytes, tumor nests, and fibroblasts. The M-Duo Technology® revealed differences in in situ cytokine (IL-6) and chemokine (CCL2, CXCL10, and IL-8) secretion between Mel-RhS and the control RhS. Notably, such differences were not detected through conventional investigation of secreted proteins in culture supernatants.The combination of LC-OCT's high-resolution imaging and M-Duo Technology®'s in situ microenvironmental mapping has the potential to provide a synergistic platform for non-invasive, real-time analysis, allowing for prolonged observation of processes within Mel-RhS models without the need for culture disruption.Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.