Development of Cell-Derived Matrices for Three-Dimensional In Vitro Cancer Cell Models

Abstract

Most morphogenetic and pathological processes are driven by cells responding to the surrounding matrix, such as its composition, architecture, and mechanical properties. Despite increasing evidence for the role of extracellular matrix (ECM) in tissue and disease development, many in vitro substitutes still fail to effectively mimic the native microenvironment. We established a novel method to produce macroscale (>1 cm) mesenchymal cell-derived matrices (CDMs) aimed to mimic the fibrotic tumor microenvironment surrounding epithelial cancer cells. CDMs are produced by human adipose mesenchymal stem cells cultured in sacrificial 3D scaffold templates of fibronectin-coated poly-lactic acid microcarriers (MCs) in the presence of macromolecular crowders. We showed that decellularized CDMs closely mimic the fibrillar protein composition, architecture, and mechanical properties of human fibrotic ECM from cancer masses. CDMs had highly reproducible composition made of collagen types I and III and fibronectin ECM with tunable mechanical properties. Moreover, decellularized and MC-free CDMs were successfully repopulated with cancer cells throughout their 3D structure, and following chemotherapeutic treatment, cancer cells showed greater doxorubicin resistance compared to 3D culture in collagen hydrogels. Collectively, these results support the use of CDMs as a reproducible and tunable tool for developing 3D in vitro cancer models.

Combining Cryogel Architecture and Macromolecular Crowding‐Enhanced Extracellular Matrix Cues to Mimic the Bone Marrow Niche

“…To our knowledge, there are only a few examples where crowding molecules have been applied in a 3D culture system. [83][84][85] To date, there are no studies involving bone marrow-derived MSCs. The effect of a mixture of Ficoll 70 and Ficoll 400 on the ECM production by porcine chondrocytes grown in both 2D cell monolayers and 3D cartilage cultures was investigated.…”

Brain Cancer Cell-Derived Matrices and Effects on Astrocyte Migration

“…Additionally, the tumor microenvironment has been proposed as a source of "pro-regenerative" -or at least anti-inflammatory -cues [Gregory and Paterson, 2018], and cancerderived ECM may help inform development of therapeutic biomaterials. Previous studies have decellularized cellderived matrices and replated new cells to show the effects on subsequent cell migration and metabolic function [Kaukonen et al, 2017;Rubí-Sans et al, 2021]. We chose to focus on decellularizing the cancer-derived matrix created using Ficoll MMC, since this condition promoted the most matrix accumulation.…”

“…Common MMC agents are high molecular weight polysaccharides -like Ficoll, carrageenan, and dextran sulfate -because a single molecule can exclude significant medium volume while also being relatively inert. Crowding has been used to increase production and/or deposition of ECM molecules in cultures of fibroblasts [Marinkovic et al, 2021;Shendi et al, 2019;Kaukonen et al, 2017], mesenchymal stem cells [Chiang et al, 2021;Rubí-Sans et al, 2021], and some cancer cell types [Bascetin et al, 2021;Ranamukhaarachchi et al, 2019]. This approach has been proposed for applications in screening of antifibrotic drugs [Cavanzo et al,2020], modeling the fibrotic tumor microenvironment [Rubí-Sans et al, 2021], and creating engineered scaffolds for tissue regeneration [Fitzpatrick and McDevitt, 2015].…”

“…Crowding has been used to increase production and/or deposition of ECM molecules in cultures of fibroblasts [Marinkovic et al, 2021;Shendi et al, 2019;Kaukonen et al, 2017], mesenchymal stem cells [Chiang et al, 2021;Rubí-Sans et al, 2021], and some cancer cell types [Bascetin et al, 2021;Ranamukhaarachchi et al, 2019]. This approach has been proposed for applications in screening of antifibrotic drugs [Cavanzo et al,2020], modeling the fibrotic tumor microenvironment [Rubí-Sans et al, 2021], and creating engineered scaffolds for tissue regeneration [Fitzpatrick and McDevitt, 2015]. MMC protocols vary widely, and the best molecule to promote ECM deposi-tion appears to be cell-specific.…”

Evaluation of Proton-Induced DNA Damage in 3D-Engineered Glioblastoma Microenvironments

“…In the field of engineered 3D cell microenvironments, the development of architectures for the study and treatment of cancer cells assumes a paramount importance for fundamental mechanobiology and prospective clinical studies. − Among the different types of cancer, glioblastoma (GBM) is a grade IV glioma brain tumor and the most aggressive type of brain cancer with an incidence of 2–3 cases per 100.000 people per year . GBM continues to have one of the most dismal prognoses of any cancer with a median survival rate of about 12–15 months and less than 5% of cases surviving over 5 years after initial diagnosis. , The current treatment regime includes removal of the bulk of the tumor by surgery followed by image-guided radiotherapy (where high-energy X-ray radiation destroys the cancer cells) and chemotherapy to control the regrowth of the tumor.…”