A Scaffold-Free 3-D Co-Culture Mimics the Major Features of the Reverse Warburg Effect In Vitro
Most tumors consume large amounts of glucose. Concepts to explain the mechanisms that mediate the achievement of this metabolic need have proposed a switch of the tumor mass to aerobic glycolysis. Depending on whether primarily tumor or stroma cells undergo such a commutation, the terms ‘Warburg eff...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-08-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/8/1900 |
| _version_ | 1797558209930592256 |
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| author | Florian Keller Roman Bruch Richard Schneider Julia Meier-Hubberten Mathias Hafner Rüdiger Rudolf |
| author_facet | Florian Keller Roman Bruch Richard Schneider Julia Meier-Hubberten Mathias Hafner Rüdiger Rudolf |
| author_sort | Florian Keller |
| collection | DOAJ |
| description | Most tumors consume large amounts of glucose. Concepts to explain the mechanisms that mediate the achievement of this metabolic need have proposed a switch of the tumor mass to aerobic glycolysis. Depending on whether primarily tumor or stroma cells undergo such a commutation, the terms ‘Warburg effect’ or ‘reverse Warburg effect’ were coined to describe the underlying biological phenomena. However, current in vitro systems relying on 2-D culture, single cell-type spheroids, or basal-membrane extract (BME/Matrigel)-containing 3-D structures do not thoroughly reflect these processes. Here, we aimed to establish a BME/Matrigel-free 3-D microarray cancer model to recapitulate the metabolic interplay between cancer and stromal cells that allows mechanistic analyses and drug testing. Human HT-29 colon cancer and CCD-1137Sk fibroblast cells were used in mono- and co-cultures as 2-D monolayers, spheroids, and in a cell-chip format. Metabolic patterns were studied with immunofluorescence and confocal microscopy. In chip-based co-cultures, HT-29 cells showed facilitated 3-D growth and increased levels of hexokinase-2, TP53-induced glycolysis and apoptosis regulator (TIGAR), lactate dehydrogenase, and: translocase of outer mitochondrial membrane 20 (TOMM20), when compared with HT-29 mono-cultures. Fibroblasts co-cultured with HT-29 cells expressed higher levels of mono-carboxylate transporter 4, hexokinase-2, microtubule-associated proteins 1A/1B light chain 3, and ubiquitin-binding protein p62 than in fibroblast mono-cultures, in both 2-D cultures and chips. Tetramethylrhodamin-methylester (TMRM) live-cell imaging of chip co-cultures revealed a higher mitochondrial potential in cancer cells than in fibroblasts. The findings demonstrate a crosstalk between cancer cells and fibroblasts that affects cellular growth and metabolism. Chip-based 3-D co-cultures of cancer cells and fibroblasts mimicked features of the reverse Warburg effect. |
| first_indexed | 2024-03-10T17:28:03Z |
| format | Article |
| id | doaj.art-1671a70e032a49b483d265d2aa33858a |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T17:28:03Z |
| publishDate | 2020-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-1671a70e032a49b483d265d2aa33858a2023-11-20T10:06:09ZengMDPI AGCells2073-44092020-08-0198190010.3390/cells9081900A Scaffold-Free 3-D Co-Culture Mimics the Major Features of the Reverse Warburg Effect In VitroFlorian Keller0Roman Bruch1Richard Schneider2Julia Meier-Hubberten3Mathias Hafner4Rüdiger Rudolf5Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences, 68163 Mannheim, GermanyInstitute of Molecular and Cell Biology, Mannheim University of Applied Sciences, 68163 Mannheim, GermanyTIP Oncology, Merck Healthcare KGaA, 64289 Darmstadt, GermanyTIP Oncology, Merck Healthcare KGaA, 64289 Darmstadt, GermanyInstitute of Molecular and Cell Biology, Mannheim University of Applied Sciences, 68163 Mannheim, GermanyInstitute of Molecular and Cell Biology, Mannheim University of Applied Sciences, 68163 Mannheim, GermanyMost tumors consume large amounts of glucose. Concepts to explain the mechanisms that mediate the achievement of this metabolic need have proposed a switch of the tumor mass to aerobic glycolysis. Depending on whether primarily tumor or stroma cells undergo such a commutation, the terms ‘Warburg effect’ or ‘reverse Warburg effect’ were coined to describe the underlying biological phenomena. However, current in vitro systems relying on 2-D culture, single cell-type spheroids, or basal-membrane extract (BME/Matrigel)-containing 3-D structures do not thoroughly reflect these processes. Here, we aimed to establish a BME/Matrigel-free 3-D microarray cancer model to recapitulate the metabolic interplay between cancer and stromal cells that allows mechanistic analyses and drug testing. Human HT-29 colon cancer and CCD-1137Sk fibroblast cells were used in mono- and co-cultures as 2-D monolayers, spheroids, and in a cell-chip format. Metabolic patterns were studied with immunofluorescence and confocal microscopy. In chip-based co-cultures, HT-29 cells showed facilitated 3-D growth and increased levels of hexokinase-2, TP53-induced glycolysis and apoptosis regulator (TIGAR), lactate dehydrogenase, and: translocase of outer mitochondrial membrane 20 (TOMM20), when compared with HT-29 mono-cultures. Fibroblasts co-cultured with HT-29 cells expressed higher levels of mono-carboxylate transporter 4, hexokinase-2, microtubule-associated proteins 1A/1B light chain 3, and ubiquitin-binding protein p62 than in fibroblast mono-cultures, in both 2-D cultures and chips. Tetramethylrhodamin-methylester (TMRM) live-cell imaging of chip co-cultures revealed a higher mitochondrial potential in cancer cells than in fibroblasts. The findings demonstrate a crosstalk between cancer cells and fibroblasts that affects cellular growth and metabolism. Chip-based 3-D co-cultures of cancer cells and fibroblasts mimicked features of the reverse Warburg effect.https://www.mdpi.com/2073-4409/9/8/1900reverse Warburg effectfibroblastsMCT4LC3mitochondriaoptical tissue clearing |
| spellingShingle | Florian Keller Roman Bruch Richard Schneider Julia Meier-Hubberten Mathias Hafner Rüdiger Rudolf A Scaffold-Free 3-D Co-Culture Mimics the Major Features of the Reverse Warburg Effect In Vitro Cells reverse Warburg effect fibroblasts MCT4 LC3 mitochondria optical tissue clearing |
| title | A Scaffold-Free 3-D Co-Culture Mimics the Major Features of the Reverse Warburg Effect In Vitro |
| title_full | A Scaffold-Free 3-D Co-Culture Mimics the Major Features of the Reverse Warburg Effect In Vitro |
| title_fullStr | A Scaffold-Free 3-D Co-Culture Mimics the Major Features of the Reverse Warburg Effect In Vitro |
| title_full_unstemmed | A Scaffold-Free 3-D Co-Culture Mimics the Major Features of the Reverse Warburg Effect In Vitro |
| title_short | A Scaffold-Free 3-D Co-Culture Mimics the Major Features of the Reverse Warburg Effect In Vitro |
| title_sort | scaffold free 3 d co culture mimics the major features of the reverse warburg effect in vitro |
| topic | reverse Warburg effect fibroblasts MCT4 LC3 mitochondria optical tissue clearing |
| url | https://www.mdpi.com/2073-4409/9/8/1900 |
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