Arcuri Sharon, Pennarossa Georgia, Prasadani Madhusha, Gandolfi Fulvio, Brevini Tiziana A L
Department of Veterinary Medicine, Università degli Studi di Sassari, Via Vienna, 07100 Sassari, Italy.
Laboratory of Biomedical Embryology and Tissue Engineering, Department of Veterinary Medicine and Animal Sciences, Università degli Studi di Milano, Via dell'Università 6, 26900 Lodi, Italy.
Methods Protoc. 2024 Sep 27;7(5):76. doi: 10.3390/mps7050076.
In recent years, great interest has been focused on the development of highly reproducible 3D in vitro models that are able to mimic the physiological architecture and functionality of native tissues. To date, a wide range of techniques have been proposed to recreate an intestinal barrier in vitro, including synthetic scaffolds and hydrogels, as well as complex on-a-chip systems and organoids. Here, we describe a novel protocol for the generation of an artificial intestine based on the creation of decellularized bio-scaffolds and their repopulation with intestinal stromal and epithelial cells. Organs collected at the local slaughterhouse are subjected to a decellularization protocol that includes a freezing/thawing step, followed by sequential incubation in 1% SDS for 12 h, 1% Triton X-100 for 12 h, and 2% deoxycholate for 12 h. At the end of the procedure, the generated bio-scaffolds are repopulated with intestinal fibroblasts and then with epithelial cells. The protocol described here represents a promising and novel strategy to generate an in vitro bioengineered intestine platform able to mimic some of the complex functions of the intestinal barrier, thus constituting a promising 3D strategy for nutritional, pharmaceutical, and toxicological studies.
近年来，人们高度关注能够模拟天然组织生理结构和功能的高度可重复的三维体外模型的开发。迄今为止，已经提出了多种技术来在体外重建肠道屏障，包括合成支架和水凝胶，以及复杂的芯片系统和类器官。在此，我们描述了一种基于脱细胞生物支架的创建及其用肠道基质细胞和上皮细胞重新填充来生成人工肠道的新方案。在当地屠宰场收集的器官要经过脱细胞处理方案，该方案包括冷冻/解冻步骤，随后依次在1%十二烷基硫酸钠（SDS）中孵育12小时、在1% Triton X-100中孵育12小时以及在2%脱氧胆酸盐中孵育12小时。在该过程结束时，生成的生物支架先用肠道成纤维细胞重新填充，然后再用上皮细胞重新填充。本文所述方案是一种有前景的新策略，用于生成能够模拟肠道屏障一些复杂功能的体外生物工程肠道平台，从而构成用于营养、制药和毒理学研究的一种有前景的三维策略。
