In Vitro 3D Cultures to Model the Tumor Microenvironment

Fontana, Fabrizio; Marzagalli, Monica; Sommariva, Michele; Gagliano, Nicoletta; Limonta, Patrizia

doi:10.3390/cancers13122970

Cited by 59 publications

(43 citation statements)

References 125 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Next, PFs and CAFs used in this study were not patientmatched, therefore using CAFs from different patient donors would have been desirable. Prospectively, it would be interesting to investigate the PDAC SiaJ in a co-culture system, as the interaction between cancer cells, immune cells, and fibroblasts is extremely important and could influence different treatment strategies [87,88]. In addition, culture cells in other ECM-specific substrates might influence cell phenotype [89].…”

Section: Discussionmentioning

confidence: 99%

Collagen Biomarkers Quantify Fibroblast Activity In Vitro and Predict Survival in Patients with Pancreatic Ductal Adenocarcinoma

Nissen

Johansen

Chen

et al. 2022

Cancers

View full text Add to dashboard Cite

The use of novel tools to understand tumour-fibrosis in pancreatic ductal adenocarcinoma (PDAC) and novel anti-fibrotic treatments are highly needed. We established a pseudo-3D in vitro model including humane pancreatic fibroblasts (PFs) and pancreatic cancer-associated fibroblasts (CAFs) in combination with clinical collagen biomarkers, as a translational anti-fibrotic drug screening tool. Furthermore, we investigated the prognostic potential of serum collagen biomarkers in 810 patients with PDAC. PFs and CAFs were cultured in Ficoll-media. Cells were treated w/wo TGF-ß1 and the anti-fibrotic compound ALK5i. Biomarkers measuring the formation of type III (PRO-C3) and VI (PRO-C6) collagens were measured by ELISA in supernatant at days 3, 6, 9, and 12. PRO-C3 and PRO-C6, and their association with overall survival (OS), were evaluated in serum with PDAC (n = 810). PRO-C3 and PRO-C6 were upregulated in CAFs compared to PFs (p < 0.0001.). TGF-ß1 increased PRO-C3 in both PFs and CAFs (p < 0.0001). The anti-fibrotic compound ALK5i inhibited both PRO-C3 and PRO-C6 (p < 0.0001). High serum levels of PRO-C3 and PRO-C6 in patients with PDAC were associated with short OS (PRO-C3:HR= 1.48, 95%CI:1.29–1.71, p < 0.0001 and PRO-C6:HR = 1.31, 95%CI:1.14–1.50, p = 0.0002). PRO-C3 and PRO-C6 have the potential to be used both pre-clinically and clinically as a measure of tumor fibrosis and CAF activity.

show abstract

Section: Discussionmentioning

confidence: 99%

Collagen Biomarkers Quantify Fibroblast Activity In Vitro and Predict Survival in Patients with Pancreatic Ductal Adenocarcinoma

Nissen

Johansen

Chen

et al. 2022

Cancers

View full text Add to dashboard Cite

show abstract

“…There are some data about the treatment sensitivity differences of these models, which highlight that 3D structures and cellular complexity could influence the experimental results of in vitro therapeutic drug sensitivity tests [51,55]. Gene expression analyses data from these 2D vs. 3D spheroid cultures described some well-known and expected alterations in glycolysis, PI3K/Akt pathways and/or ECM protein and other metabolic events (e.g., mitophagy) and highlighted the importance of metabolic rewiring differences in the used in vitro models [51,55,56]. The cellular and ECM heterogeneity could have particular significance in immune-oncology drug tests and their in vitro experiments in future studies, as several papers suggested [52,53].…”

Section: Discussionmentioning

confidence: 94%

Characterisation of 3D Bioprinted Human Breast Cancer Model for In Vitro Drug and Metabolic Targeting

Dankó

Petővári

Raffay

et al. 2022

IJMS

View full text Add to dashboard Cite

Monolayer cultures, the less standard three-dimensional (3D) culturing systems, and xenografts are the main tools used in current basic and drug development studies of cancer research. The aim of biofabrication is to design and construct a more representative in vivo 3D environment, replacing two-dimensional (2D) cell cultures. Here, we aim to provide a complex comparative analysis of 2D and 3D spheroid culturing, and 3D bioprinted and xenografted breast cancer models. We established a protocol to produce alginate-based hydrogel bioink for 3D bioprinting and the long-term culturing of tumour cells in vitro. Cell proliferation and tumourigenicity were assessed with various tests. Additionally, the results of rapamycin, doxycycline and doxorubicin monotreatments and combinations were also compared. The sensitivity and protein expression profile of 3D bioprinted tissue-mimetic scaffolds showed the highest similarity to the less drug-sensitive xenograft models. Several metabolic protein expressions were examined, and the in situ tissue heterogeneity representing the characteristics of human breast cancers was also verified in 3D bioprinted and cultured tissue-mimetic structures. Our results provide additional steps in the direction of representing in vivo 3D situations in in vitro studies. Future use of these models could help to reduce the number of animal experiments and increase the success rate of clinical phase trials.

show abstract

“…Interestingly, we found that TAM heterogeneity is lost when isolated and cultured ex vivo in 2D conditions, emphasizing the need for TME signals to maintain their identity. In recent years, 3D cultures in hydrogels, including collagen [ 55 , 56 ], have provided valuable tools to accelerate tumor studies and TME modeling [ 33 , 34 ]. In the present study, we developed a 3D collagen co-culture system to mimic the melanoma TME and investigate how interactions between melanoma cells, fibroblasts, and macrophages shape the early stages of macrophage immune activity leading to such complex phenotypes.…”

Section: Discussionmentioning

confidence: 99%

“…Recent studies have demonstrated that organotypic melanoma cultures outperform 2D assays when studying TME-imprinting mechanisms and closely resemble tumor growth as observed in human lesions while supporting cell survival and function [ 32 , 33 ]. The use of 3D cultures may accelerate the identification of predictive and/or prognostic markers and the development of effective combination therapies [ 34 ]. Ex vivo systems that incorporate key features of the native TME and model the dynamic response to checkpoint inhibitors can facilitate efforts in precision immuno-oncology.…”

Section: Introductionmentioning

confidence: 99%

3D Model of the Early Melanoma Microenvironment Captures Macrophage Transition into a Tumor-Promoting Phenotype

Pizzurro

Liu

Bridges

et al. 2021

Cancers

View full text Add to dashboard Cite

Tumor immune response is shaped by the tumor microenvironment (TME), which often evolves to be immunosuppressive, promoting disease progression and metastasis. An important example is melanoma tumors, which display high numbers of tumor-associated macrophages (TAMs) that are immunosuppressive but also have the potential to restore anti-tumor activity. However, to therapeutically target TAMs, there is a need to understand the early events that shape their tumor-promoting profile. To address this, we built and optimized 3D in vitro co-culture systems, composed of a collagen-I matrix scaffolding murine bone-marrow-derived macrophages (BMDMs), YUMM1.7 melanoma cells, and fibroblasts to recreate the early melanoma TME and study how interactions with fibroblasts and tumor cells modulate macrophage immune activity. We monitored BMDM behavior and interactions through time-lapse imaging and characterized their activation and secretion. We found that stromal cells induced a rapid functional activation, with increased motility and response from BMDMs. Over the course of seven days, BMDMs acquired a phenotype and secretion profile that resembled melanoma TAMs in established tumors. Overall, the direct cell–cell interactions with the stromal components in a 3D environment shape BMDM transition to a TAM-like immunosuppressive state. Our systems will enable future studies of changes in macrophage–stromal cross-talk in the melanoma TME

show abstract

In Vitro 3D Cultures to Model the Tumor Microenvironment

Cited by 59 publications

References 125 publications

Collagen Biomarkers Quantify Fibroblast Activity In Vitro and Predict Survival in Patients with Pancreatic Ductal Adenocarcinoma

Collagen Biomarkers Quantify Fibroblast Activity In Vitro and Predict Survival in Patients with Pancreatic Ductal Adenocarcinoma

Characterisation of 3D Bioprinted Human Breast Cancer Model for In Vitro Drug and Metabolic Targeting

3D Model of the Early Melanoma Microenvironment Captures Macrophage Transition into a Tumor-Promoting Phenotype

Contact Info

Product

Resources

About