Three-dimensional culture suppresses pancreatic fibroblast activation and limits paracrine signaling between pancreatic cancer cells and fibroblasts
- 1Department of Visceral, Vascular and Endocrine Surgery, Martin-Luther-University Halle-Wittenberg, University Medical Center Halle, Halle (Saale), 06120, Germany
- 2Institute of Medical and Public Health Research, Ilia State University, Tbilisi, 0162, Georgia
Abstract
Background: Pancreatic fibroblasts play a significant role in the development, progression, and treatment limitations of pancreatic ductal adenocarcinoma (PDAC), making them an essential area of research. Traditional 2D in vitro models fail to fully capture complexities of these stromal cells in PDAC such as interactions, morphology, and response to stimuli present in in vivo environments, making 3D culture systems a promising platform for further studies.
Methods: We compared the behavior of pancreatic fibroblasts in 2D and 3D culture systems, by analyzing the effects of different cultures on transcriptomics. Primary murine pancreatic fibroblasts were cultured in monolayers and 3D spheroids. An indirect co-culture system between fibroblasts and KC murine pancreatic cancer cells (KCs) served to investigate paracrine effects of cancer cells. RNA sequencing and RT-PCR were utilized for data analysis, supported by Oil Red O staining.
Results: Inactivation of fibroblasts in the 3D magnetic levitation model was noted by downregulation of activation markers Acta2, Fap, and Ctgf, and upregulation of a lipid-droplet stabilizer Plin2. The HIF-1 signaling pathway was enriched in the 3D model compared to 2D. Oil Red O staining demonstrated upregulation of cytoplasmic lipid droplets, in 3D vs. 2D culture models. Activation marker downregulation and lipid accumulation seen in both monocultures and co-cultures in 3D indicated a cell-specific behavior rather than tumor cell influence.
Conclusion: The 3D magnetic levitation culture could serve as a model to study fibroblasts in an inactivated state, while co-culture with KC tumor cells could be helpful for cancer-fibroblast paracrine interaction studies.
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BibTeX
@article{javakhishvili2026threedimensional,
title = {{Three-dimensional culture suppresses pancreatic fibroblast activation and limits paracrine signaling between pancreatic cancer cells and fibroblasts}},
author = {Nino Javakhishvili and Anais Zourelidis and Bogusz Trojanowicz and Yoshiaki Sunami and Jörg Kleeff},
journal = {Biochemistry and Biophysics Reports},
year = {2026},
month = oct,
doi = {10.1016/j.bbrep.2026.102843},
url = {https://doi.org/10.1016/j.bbrep.2026.102843},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Three-dimensional culture suppresses pancreatic fibroblast activation and limits paracrine signaling between pancreatic cancer cells and fibroblasts
AU - Nino Javakhishvili
AU - Anais Zourelidis
AU - Bogusz Trojanowicz
AU - Yoshiaki Sunami
AU - Jörg Kleeff
PY - 2026
DA - 2026/10/27
JO - Biochemistry and Biophysics Reports
DO - 10.1016/j.bbrep.2026.102843
UR - https://doi.org/10.1016/j.bbrep.2026.102843
AB - Background: Pancreatic fibroblasts play a significant role in the development, progression, and treatment limitations of pancreatic ductal adenocarcinoma (PDAC), making them an essential area of research. Traditional 2D in vitro models fail to fully capture complexities of these stromal cells in PDAC such as interactions, morphology, and response to stimuli present in in vivo environments, making 3D culture systems a promising platform for further studies. Methods: We compared the behavior of pancreatic fibroblasts in 2D and 3D culture systems, by analyzing the effects of different cultures on transcriptomics. Primary murine pancreatic fibroblasts were cultured in monolayers and 3D spheroids. An indirect co-culture system between fibroblasts and KC murine pancreatic cancer cells (KCs) served to investigate paracrine effects of cancer cells. RNA sequencing and RT-PCR were utilized for data analysis, supported by Oil Red O staining. Results: Inactivation of fibroblasts in the 3D magnetic levitation model was noted by downregulation of activation markers Acta2, Fap, and Ctgf, and upregulation of a lipid-droplet stabilizer Plin2. The HIF-1 signaling pathway was enriched in the 3D model compared to 2D. Oil Red O staining demonstrated upregulation of cytoplasmic lipid droplets, in 3D vs. 2D culture models. Activation marker downregulation and lipid accumulation seen in both monocultures and co-cultures in 3D indicated a cell-specific behavior rather than tumor cell influence. Conclusion: The 3D magnetic levitation culture could serve as a model to study fibroblasts in an inactivated state, while co-culture with KC tumor cells could be helpful for cancer-fibroblast paracrine interaction studies.
SN - 2405-5808
ER -