Jul 2026· Journal of Advanced Research· 0 citations· 85 references
Medicine
TL;DR
The 3D co-culture model provides a biomimetic platform for simulating the PCa bone-metastatic microenvironment and may serve as a useful tool for preclinical drug screening and evaluation of combination treatment strategies for bone-metastatic prostate cancer.
Abstract
INTRODUCTION
Prostate cancer (PCa) is one of the most prevalent malignancies in men and frequently progresses to bone metastasis. Understanding the interactions between PCa cells and the bone microenvironment, as well as their impact on therapeutic response, is therefore of critical clinical relevance.
Objectives
This study aimed to investigate the effects of the osteogenic microenvironment on prostate cancer cells by constructing three-dimensional (3D) bioprinted in vitro co-culture models, and to evaluate how the osteogenic niche influences tumor malignant phenotypes.
Methods
This study employed extrusion-based 3D bioprinting (3DP) to construct in vitro co-culture models of the PCa osteogenic microenvironment. Two 3D-PCa models based on LNCaP and PC-3 cells were each co-cultured with osteogenically differentiated adipose-derived stem cells (ADSCs) to generate corresponding osteogenic niche models. Tumor phenotypes and drug responses were evaluated using functional assays, histological and immunofluorescence analyses, molecular profiling, and RNA sequencing.
Results
The 3D bioprinted constructs exhibited structural stability and high reproducibility, providing a 3D growth environment that mimics key aspects of the in vivo tumor niche. The 3D-PCa models showed enhanced drug resistance, invasive potential, and adaptation to androgen-deprivation. Meanwhile, ADSCs exhibited robust osteogenic differentiation within 3D scaffolds. Under co-culture conditions, 3D-PCa cells exhibited enhanced malignant-like phenotypes, including increased proliferation and reduced drug sensitivity. Multi-level analyses further indicated that these phenotypic changes were associated with coordinated activation of epithelial-mesenchymal transition (EMT)-, hypoxia-, and mitogen-activated protein kinase (MAPK)-related signaling programs, accompanied by alterations in drug transporter expression and cell cycle distribution.
Conclusion
The 3D co-culture model provides a biomimetic platform for simulating the PCa bone-metastatic microenvironment. This system enables investigation of the effects of the osteogenic niche on tumor behavior and therapeutic responses, and may serve as a useful tool for preclinical drug screening and evaluation of combination treatment strategies for bone-metastatic prostate cancer.
In breast cancer, local invasion of cancer cells into surrounding tissue marks the first step of metastasis. However, to elucidate the impact of cells from the tumor microenvironment on this process, advanced 3D migration models are still urgently needed. To enable migration and invasion studies in a fully 3D bioprinted tumor‐stroma model, a migration‐permissive bioink composed of methacrylated collagen type I and thiolated hyaluronic acid with low polymer content is developed. In a printed co‐culture model comprising metastatic breast cancer cells (MDA‐MB‐231) and adipose‐derived stromal cells (ASCs), real‐time single‐cell tracking reveals that ASCs in the stromal compartment profoundly promote migration and invasion dynamics of individual tumor cells. This is reflected by increased speed, migration distance, and invasion into the stroma, and is accompanied by collagen remodeling and a shift in tumor cell morphology. A correlation between tumor cell morphology and migration speed is evident, which is modulated by ASCs. A highly motile and invasive subset of tumor cells is significantly enhanced in the presence of ASCs. These insights into the influence of ASCs on the heterogeneity of breast cancer cells in terms of their migratory behavior may inform the development of more specific and effective treatment options for metastatic breast cancer.
Sabrina Stecher, Joachim Schenk, Alessandro Cianciosi et al.· Advanced Healthcare Material...· 0 citations
One of the major challenges in late-stage cancer research is studying the direct interactions between cancer cells and the metastatic tissue. Metastasis is an inefficient multistep process in which only a small subset of cancer cells successfully colonizes the new microenvironment. Bone is the most common tissue for breast and prostate cancer metastasis and the third most common site across all cancer types. We present an ex vivo co-culture protocol for modeling bone metastasis that enables direct investigation of cancer cell interactions with the native bone and marrow microenvironment. The procedure involves the extraction and preparation of mouse long bones (femurs and tibiae), injection of cancer cells into the bone marrow cavity, maintenance of the co-culture, and sample processing for downstream analysis. The protocol supports functional readouts, including bioluminescence imaging of tumor cell viability, conditioned media sampling, and downstream tissue analysis, such as immunohistochemistry or micro-computed tomography (µCT). The culture setup procedure can be completed in a single day, and daily media maintenance thereafter requires minimal time. Final sample processing times vary, depending on the analysis completed. The system maintains viable and metabolically active bone, marrow, and cancer cells for up to four weeks and preserves the heterogeneity of the bone microenvironment. It is highly adaptable for various cancer types and bone genotypes and enables high-throughput functional interrogation of cancer colonization in a complex microenvironment. The protocol requires basic expertise in mammalian cell culture and mouse dissection and is suitable for researchers studying metastasis, tumor–stroma interactions, or preclinical therapeutic screening in the bone microenvironment. Not applicable.
Ricardo Romero-Moreno, Courtney L. Flatt, Madeline P. Sheeley et al.· BMC Methods· 0 citations
Optization of MC-based 3D culture conditions showed that medium viscosity affects both spheroid morphology and transcriptional regulation, indicating that the physical properties of the microenvironment influence ESR1 expression and ERα activity.
Gozde Korkmaz, Elif Kansız· Istanbul University Journal...· 0 citations
Glioblastoma (GBM) is the most aggressive primary brain tumor and remains associated with poor clinical outcomes despite advances in surgical and adjuvant therapies. The tumor microenvironment, particularly extracellular matrix (ECM) interactions, plays a crucial role in regulating glioblastoma progression, cellular plasticity, and therapeutic resistance. Therefore, physiologically relevant three-dimensional (3D) models are needed to better recapitulate GBM biology. In this study, we investigated the effects of ECM-functionalized polyacrylonitrile/coumarin-500 (PAN/C500) nanofiber scaffolds on the phenotype of LN-18 and U-87 MG glioblastoma cells cultured under 3D conditions. Cytoskeletal organization was assessed by phalloidin staining and live-cell vimentin imaging, while epithelial–mesenchymal transition (EMT)-associated proteins and stemness-related markers were analyzed by Western blotting. ECM-functionalized 3D PAN/C500 scaffolds promoted significant cytoskeletal remodeling, altered EMT-associated protein expression, and increased the expression of stemness-associated proteins, particularly SOX2, NANOG, and Nestin, compared with conventional 2D cultures. These responses were accompanied by cell line-dependent phenotypic adaptations, indicating that the engineered microenvironment influences glioblastoma cell behavior. This platform may serve as a valuable model for investigating glioblastoma biology and microenvironment-associated molecular adaptations in vitro.
Ihsan Nalkiran, Hatice Sevim Nalkıran, Derya Bal Altuntaş et al.· Biomolecules· 0 citations