Bilayer 3D co‐culture platform inducing the differentiation of normal fibroblasts into cancer‐associated fibroblast like cells: New in vitro source to obtain cancer‐associated fibroblasts
Abstract This study presents a novel in vitro bilayer 3D co‐culture platform designed to obtain cancer‐associated fibroblasts (CAFs)‐like cells. The platform consists of a bilayer hydrogel structure with a collagen/polyethylene glycol (PEG) hydrogel for fibroblasts as the upper layer and an alginate...
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| Format: | Article |
| Language: | English |
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Wiley
2025-01-01
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| Series: | Bioengineering & Translational Medicine |
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| Online Access: | https://doi.org/10.1002/btm2.10708 |
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| author | Yeon Ju Kim Hyeon Song Lee Dohyun Kim Hwa Kyung Byun Woong Sub Koom Won‐Gun Koh |
| author_facet | Yeon Ju Kim Hyeon Song Lee Dohyun Kim Hwa Kyung Byun Woong Sub Koom Won‐Gun Koh |
| author_sort | Yeon Ju Kim |
| collection | DOAJ |
| description | Abstract This study presents a novel in vitro bilayer 3D co‐culture platform designed to obtain cancer‐associated fibroblasts (CAFs)‐like cells. The platform consists of a bilayer hydrogel structure with a collagen/polyethylene glycol (PEG) hydrogel for fibroblasts as the upper layer and an alginate hydrogel for tumor cells as the lower layer. The platform enabled paracrine interactions between fibroblasts and cancer cells, which allowed for selective retrieval of activated fibroblasts through collagenase treatment for further study. Fibroblasts remained viable throughout the culture periods and showed enhanced proliferation when co‐cultured with cancer cells. Morphological changes in the co‐cultured fibroblasts resembling CAFs were observed, especially in the 3D microenvironment. The mRNA expression levels of CAF‐related markers were significantly upregulated in 3D, but not in 2D co‐culture. Proteomic analysis identified upregulated proteins associated with CAFs, further confirming the transformation of normal fibroblasts into CAF within the proposed 3D co‐culture platform. Moreover, co‐culture with CAF induced radio‐ and chemoresistance in pancreatic cancer cells (PANC‐1). Survival rate of cancer cells post‐irradiation and gemcitabine resistance increased significantly in the co‐culture setting, highlighting the role of CAFs in promoting cancer cell survival and therapeutic resistance. These findings would contribute to understanding molecular and phenotypic changes associated with CAF activation and provide insights into potential therapeutic strategies targeting the tumor microenvironment. |
| format | Article |
| id | doaj-art-83d5d683c9444bc083dc03ee5d6f1f38 |
| institution | Kabale University |
| issn | 2380-6761 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Bioengineering & Translational Medicine |
| spelling | doaj-art-83d5d683c9444bc083dc03ee5d6f1f382025-01-09T06:19:46ZengWileyBioengineering & Translational Medicine2380-67612025-01-01101n/an/a10.1002/btm2.10708Bilayer 3D co‐culture platform inducing the differentiation of normal fibroblasts into cancer‐associated fibroblast like cells: New in vitro source to obtain cancer‐associated fibroblastsYeon Ju Kim0Hyeon Song Lee1Dohyun Kim2Hwa Kyung Byun3Woong Sub Koom4Won‐Gun Koh5Department of Radiation Oncology, Yonsei Cancer Center Yonsei University College of Medicine Seoul South KoreaDepartment of Chemical and Biomolecular Engineering Yonsei University Seoul South KoreaDepartment of Chemical and Biomolecular Engineering Yonsei University Seoul South KoreaDepartment of Radiation Oncology, Yongin Severance Hospital Yonsei University College of Medicine Yongin South KoreaDepartment of Radiation Oncology, Yonsei Cancer Center Yonsei University College of Medicine Seoul South KoreaDepartment of Chemical and Biomolecular Engineering Yonsei University Seoul South KoreaAbstract This study presents a novel in vitro bilayer 3D co‐culture platform designed to obtain cancer‐associated fibroblasts (CAFs)‐like cells. The platform consists of a bilayer hydrogel structure with a collagen/polyethylene glycol (PEG) hydrogel for fibroblasts as the upper layer and an alginate hydrogel for tumor cells as the lower layer. The platform enabled paracrine interactions between fibroblasts and cancer cells, which allowed for selective retrieval of activated fibroblasts through collagenase treatment for further study. Fibroblasts remained viable throughout the culture periods and showed enhanced proliferation when co‐cultured with cancer cells. Morphological changes in the co‐cultured fibroblasts resembling CAFs were observed, especially in the 3D microenvironment. The mRNA expression levels of CAF‐related markers were significantly upregulated in 3D, but not in 2D co‐culture. Proteomic analysis identified upregulated proteins associated with CAFs, further confirming the transformation of normal fibroblasts into CAF within the proposed 3D co‐culture platform. Moreover, co‐culture with CAF induced radio‐ and chemoresistance in pancreatic cancer cells (PANC‐1). Survival rate of cancer cells post‐irradiation and gemcitabine resistance increased significantly in the co‐culture setting, highlighting the role of CAFs in promoting cancer cell survival and therapeutic resistance. These findings would contribute to understanding molecular and phenotypic changes associated with CAF activation and provide insights into potential therapeutic strategies targeting the tumor microenvironment.https://doi.org/10.1002/btm2.107083D co‐culturebilayer hydrogelcancer‐associated fibroblastsfibroblast activationradio‐ and chemoresistance |
| spellingShingle | Yeon Ju Kim Hyeon Song Lee Dohyun Kim Hwa Kyung Byun Woong Sub Koom Won‐Gun Koh Bilayer 3D co‐culture platform inducing the differentiation of normal fibroblasts into cancer‐associated fibroblast like cells: New in vitro source to obtain cancer‐associated fibroblasts Bioengineering & Translational Medicine 3D co‐culture bilayer hydrogel cancer‐associated fibroblasts fibroblast activation radio‐ and chemoresistance |
| title | Bilayer 3D co‐culture platform inducing the differentiation of normal fibroblasts into cancer‐associated fibroblast like cells: New in vitro source to obtain cancer‐associated fibroblasts |
| title_full | Bilayer 3D co‐culture platform inducing the differentiation of normal fibroblasts into cancer‐associated fibroblast like cells: New in vitro source to obtain cancer‐associated fibroblasts |
| title_fullStr | Bilayer 3D co‐culture platform inducing the differentiation of normal fibroblasts into cancer‐associated fibroblast like cells: New in vitro source to obtain cancer‐associated fibroblasts |
| title_full_unstemmed | Bilayer 3D co‐culture platform inducing the differentiation of normal fibroblasts into cancer‐associated fibroblast like cells: New in vitro source to obtain cancer‐associated fibroblasts |
| title_short | Bilayer 3D co‐culture platform inducing the differentiation of normal fibroblasts into cancer‐associated fibroblast like cells: New in vitro source to obtain cancer‐associated fibroblasts |
| title_sort | bilayer 3d co culture platform inducing the differentiation of normal fibroblasts into cancer associated fibroblast like cells new in vitro source to obtain cancer associated fibroblasts |
| topic | 3D co‐culture bilayer hydrogel cancer‐associated fibroblasts fibroblast activation radio‐ and chemoresistance |
| url | https://doi.org/10.1002/btm2.10708 |
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