Chitosan-hyaluronan based 3D co-culture platform for studying the crosstalk of lung cancer cells and mesenchymal stem cells.
The controversial roles of mesenchymal stem cells (MSCs) in lung cancer development are not yet resolved because of the lack of an extracellular environment that mimics the tumor microenvironment. Three-dimensional (3D) culture system is an emerging research tool for biomedical applications such as drug screening. In this study, MSCs and human non-small cell lung carcinoma cells (A549) were co-cultured on a thin biomaterial-based substratum (hyaluronan-grafted chitosan, CS-HA; ∼2μm), and they were self-organized into the 3D tumor co-spheroids with core-shell structure. The gene expression levels of tumorigenicity markers in cancer cells associated with cancer stemness, epithelial-mesenchymal transition (EMT) property, and cell mobility were up-regulated for more than twofold in the MSC-tumor co-spheroids, through the promoted expression of certain tumor enhancers and the direct cell-cell interaction. To verify the different extents of tumorigenicity, A549 cells or those co-cultured with MSCs were transplanted into zebrafish embryos for evaluation in vivo. The tumorigenicity obtained from the zebrafish xenotransplantation model was consistent with that observed in vitro. These evidences suggest that the CS-HA substrate-based 3D co-culture platform for cancer cells and MSCs may be a convenient tool for studying the cell-cell interaction in a tumor-like microenvironment and potentially for cancer drug testing.



Mesenchymal stem cells (MSCs) have been found in several types of tumor tissues. However, the controversial roles of MSCs in cancer development are still unsolved. Chitosan and hyaluronan are commonly used materials in the biomedical field. In the current study, we co-cultured lung cancer cells and MSCs on the planar hyaluronan-grafted chitosan (CS-HA) hybrid substrates, and discovered that lung cancer cells and MSCs were rapidly self-assembled into 3D tumor spheroids with core-shell structure on the substrates after only two days in culture. Therefore, CS-HA based 3D co-culture platform can be applied to exploration of the relationship between cancer cells and MSCs and other cancer-related medical applications such as drug screening.
Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan.
Hao-Wei
HW
Han
Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan; Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan; Research and Development Center for Medical Devices, National Taiwan University, Taipei, Taiwan. Electronic address: shhsu@ntu.edu.tw.
Shan-Hui
SH
Hsu
D000072283:A549 Cells; D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D063646:Carcinogenesis / Q000473:pathology; D002452:Cell Count; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics; D048429:Cell Size / Q000187:drug effects; D048271:Chitosan / Q000494:pharmacology*; D018920:Coculture Techniques / Q000379:methods*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006820:Hyaluronic Acid / Q000494:pharmacology*; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000473:pathology; D015203:Reproducibility of Results; D018874:Spheroids, Cellular / Q000187:drug effects / Q000473:pathology; D023041:Xenograft Model Antitumor Assays; D015027:Zebrafish
D014408:Biomarkers, Tumor; D017077:Culture Media, Conditioned; D006820:Hyaluronic Acid; D048271:Chitosan
3D culture; Chitosan-hyaluronan; Lung cancer; Mesenchymal stem cells; Zebrafish transplantation