Modeling Liver Organogenesis by Recreating Three-Dimensional Collective Cell Migration: A Role for TGFβ Pathway
Three-dimensional (3D) collective cell migration (CCM) is critical for improving liver cell therapies, eliciting mechanisms of liver disease, and modeling human liver development and organogenesis. Mechanisms of CCM differ in 2D vs. 3D systems, and existing models are limited to 2D or transwell-base...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-06-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2021.621286/full |
| _version_ | 1819103910680330240 |
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| author | Ogechi Ogoke Osama Yousef Cortney Ott Allison Kalinousky Wayne Lin Claire Shamul Shatoni Ross Natesh Parashurama Natesh Parashurama Natesh Parashurama |
| author_facet | Ogechi Ogoke Osama Yousef Cortney Ott Allison Kalinousky Wayne Lin Claire Shamul Shatoni Ross Natesh Parashurama Natesh Parashurama Natesh Parashurama |
| author_sort | Ogechi Ogoke |
| collection | DOAJ |
| description | Three-dimensional (3D) collective cell migration (CCM) is critical for improving liver cell therapies, eliciting mechanisms of liver disease, and modeling human liver development and organogenesis. Mechanisms of CCM differ in 2D vs. 3D systems, and existing models are limited to 2D or transwell-based systems, suggesting there is a need for improved 3D models of CCM. To recreate liver 3D CCM, we engineered in vitro 3D models based upon a morphogenetic transition that occurs during liver organogenesis, which occurs rapidly between E8.5 and E9.5 (mouse). During this morphogenetic transition, 3D CCM exhibits co-migration (multiple cell types), thick-strand interactions with surrounding septum transversum mesenchyme (STM), branching morphogenesis, and 3D interstitial migration. Here, we engineer several 3D in vitro culture systems, each of which mimics one of these processes in vitro. In mixed spheroids bearing both liver cells and uniquely MRC-5 (fetal lung) fibroblasts, we observed evidence of co-migration, and a significant increase in length and number of liver spheroid protrusions, which was highly sensitive to transforming growth factor beta 1 (TGFβ1) stimulation. In MRC-5-conditioned medium (M-CM) experiments, we observed dose-dependent branching morphogenesis associated with an upregulation of Twist1, which was inhibited by a broad TGFβ inhibitor. In models in which liver spheroids and MRC-5 spheroids were co-cultured, we observed complex strand morphogenesis, whereby thin, linear, 3D liver cell strands attach to the MRC-5 spheroid, anchor and thicken to form permanent and thick anchoring contacts between the two spheroids. In these spheroid co-culture models, we also observed spheroid fusion and strong evidence for interstitial migration. In conclusion, we present several novel cultivation systems that recreate distinct features of liver 3D CCM. These methodologies will greatly improve our molecular, cellular, and tissue-scale understanding of liver organogenesis, liver diseases like cancer, and liver cell therapy, and will also serve as a tool to bridge conventional 2D studies and preclinical in vivo studies. |
| first_indexed | 2024-12-22T01:57:58Z |
| format | Article |
| id | doaj.art-3ff7fce8728241338f9c547f46811eec |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-22T01:57:58Z |
| publishDate | 2021-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-3ff7fce8728241338f9c547f46811eec2022-12-21T18:42:44ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852021-06-01910.3389/fbioe.2021.621286621286Modeling Liver Organogenesis by Recreating Three-Dimensional Collective Cell Migration: A Role for TGFβ PathwayOgechi Ogoke0Osama Yousef1Cortney Ott2Allison Kalinousky3Wayne Lin4Claire Shamul5Shatoni Ross6Natesh Parashurama7Natesh Parashurama8Natesh Parashurama9Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, NY, United StatesDepartment of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, NY, United StatesDepartment of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, NY, United StatesDepartment of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, NY, United StatesDepartment of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, NY, United StatesDepartment of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, NY, United StatesDepartment of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, NY, United StatesDepartment of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, NY, United StatesDepartment of Biomedical Engineering, University at Buffalo (State University of New York), Buffalo, NY, United StatesClinical and Translational Research Center, University at Buffalo (State University of New York), Buffalo, NY, United StatesThree-dimensional (3D) collective cell migration (CCM) is critical for improving liver cell therapies, eliciting mechanisms of liver disease, and modeling human liver development and organogenesis. Mechanisms of CCM differ in 2D vs. 3D systems, and existing models are limited to 2D or transwell-based systems, suggesting there is a need for improved 3D models of CCM. To recreate liver 3D CCM, we engineered in vitro 3D models based upon a morphogenetic transition that occurs during liver organogenesis, which occurs rapidly between E8.5 and E9.5 (mouse). During this morphogenetic transition, 3D CCM exhibits co-migration (multiple cell types), thick-strand interactions with surrounding septum transversum mesenchyme (STM), branching morphogenesis, and 3D interstitial migration. Here, we engineer several 3D in vitro culture systems, each of which mimics one of these processes in vitro. In mixed spheroids bearing both liver cells and uniquely MRC-5 (fetal lung) fibroblasts, we observed evidence of co-migration, and a significant increase in length and number of liver spheroid protrusions, which was highly sensitive to transforming growth factor beta 1 (TGFβ1) stimulation. In MRC-5-conditioned medium (M-CM) experiments, we observed dose-dependent branching morphogenesis associated with an upregulation of Twist1, which was inhibited by a broad TGFβ inhibitor. In models in which liver spheroids and MRC-5 spheroids were co-cultured, we observed complex strand morphogenesis, whereby thin, linear, 3D liver cell strands attach to the MRC-5 spheroid, anchor and thicken to form permanent and thick anchoring contacts between the two spheroids. In these spheroid co-culture models, we also observed spheroid fusion and strong evidence for interstitial migration. In conclusion, we present several novel cultivation systems that recreate distinct features of liver 3D CCM. These methodologies will greatly improve our molecular, cellular, and tissue-scale understanding of liver organogenesis, liver diseases like cancer, and liver cell therapy, and will also serve as a tool to bridge conventional 2D studies and preclinical in vivo studies.https://www.frontiersin.org/articles/10.3389/fbioe.2021.621286/fullcell migrationhepatocyte migrationhepatic cordsliver regenerative medicinecancer metastasisliver cancer |
| spellingShingle | Ogechi Ogoke Osama Yousef Cortney Ott Allison Kalinousky Wayne Lin Claire Shamul Shatoni Ross Natesh Parashurama Natesh Parashurama Natesh Parashurama Modeling Liver Organogenesis by Recreating Three-Dimensional Collective Cell Migration: A Role for TGFβ Pathway Frontiers in Bioengineering and Biotechnology cell migration hepatocyte migration hepatic cords liver regenerative medicine cancer metastasis liver cancer |
| title | Modeling Liver Organogenesis by Recreating Three-Dimensional Collective Cell Migration: A Role for TGFβ Pathway |
| title_full | Modeling Liver Organogenesis by Recreating Three-Dimensional Collective Cell Migration: A Role for TGFβ Pathway |
| title_fullStr | Modeling Liver Organogenesis by Recreating Three-Dimensional Collective Cell Migration: A Role for TGFβ Pathway |
| title_full_unstemmed | Modeling Liver Organogenesis by Recreating Three-Dimensional Collective Cell Migration: A Role for TGFβ Pathway |
| title_short | Modeling Liver Organogenesis by Recreating Three-Dimensional Collective Cell Migration: A Role for TGFβ Pathway |
| title_sort | modeling liver organogenesis by recreating three dimensional collective cell migration a role for tgfβ pathway |
| topic | cell migration hepatocyte migration hepatic cords liver regenerative medicine cancer metastasis liver cancer |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2021.621286/full |
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