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. 2024 Jul 4;11(32):2405084. doi: 10.1002/advs.202405084

Figure 5.

Figure 5

Preparation and Applications of dECM. The procedure to obtain dECM is described in the upper box, comprising the main steps of available protocols A–D). A) Following surgical resection, fresh colon tissues are promptly transferred to the collection medium and processed. B) In a petri dish, fat and blood clots are removed from tissues and rinsed in sterile PBS. C) Samples are cut and decellularization is achieved through chemical, enzymatic and/or physical treatments.[ 108 , 109 , 112 ] Upon completion of the process, tissue turns translucent and DNA content is significantly reduced. Immunofluorescence images in panel C show the loss of cellular content in both decellularized healthy mucosa and CRC samples (DAPI staining‐blue) while preserving the matrix structure (Laminin staining‐green). Adapted with permission.[ 111 ] Copyright 2020, MDPI. D) Finally, the decellularized scaffold can be utilized for recellularization or pulverized into a fine powder and used as hydrogel in 3D co‐culture, microfluidic devices and bioprinting.[ 102 , 110 , 112 ] The lower box shows the main applications of recellularized scaffolds. Recellularization of dECM derived from healthy mucosa, CRC or metastatic sites enables the creation of models for tumor progression and metastasis as well as in vitro platforms for drug screening or gene expression profiling.[ 109 , 110 , 111 ] E) Example of histological characterization of CRC‐derived scaffold recellularized with HT‐29 cells, hematoxylin and eosin (H&E); periodic acid‐Shiff (PAS); collagen IV staining (Col IV); IF staining with epithelial marker E‐Cadherin (E‐Cad), basement membrane marker Laminin (Lam) and DAPI [adapted from reference 111]. G) Example of dECM application in chemotherapy response studies, IC50 values indicate higher resistance to 5FU in HT29 cultured within 3D scaffolds compared to 2D monolayers. Adapted with permission.[ 111 ] Copyright 2020, MDPI. Part of the figure is modified from Servier Medical Art (http://smart.servier.com/), licensed under a Creative Common Attribution 3.0 Generic License.