Culture Shock! Investigating the Importance of Physiological Tumour-Like Conditions on Cultured Cancer Cells
Notice bibliographique
Résumé
The tumour microenvironment (TME) is complex and dynamic, characterized by poor vascularization, limited nutrient availability, hypoxia, and an acidic pH. This environment plays a critical role in cancer progression, driving hallmark changes in cancer cell metabolism, morphology, proliferation, and motility. Although mammalian cell culture is a foundational tool for investigating cancer cell biology in vitro, standard cell culture practices fall short of replicating the in vivo conditions of tumours. Recently, ‘physiologic’ cell culture media that closely resemble human plasma (e.g., Plasmax, HPLM) have been developed, along with more frequent adoption of physiological oxygen conditions (2-8% O2) rather than standard atmosphere (~18% O2). Although an improvement, these conditions are not representative of all in vivo cellular environments, particularly that of the TME. Considering the tight regulatory capacity of cancer cells to sense and respond to changes in metabolic availability, we hypothesized that alterations in nutrient and oxygen availability, even if within the physiological range, may subtly rewire the metabolism in cultured cancer cells. This may necessitate further refinement of cell-specific physiologic culture conditions to better maintain the in vivo biology of cells. In this study, we directly address this hypothesis by describing the development of a tumour microenvironment-like medium (TMEM) based on metabolomic profiling of murine pancreatic ductal adenocarcinoma tumour interstitial fluid from relevant literature. We found that murine pancreatic ductal adenocarcinoma (KPCY) cells cultured under TME-like conditions (TMEM, pH 7.0, 1.5% O2) exhibited increased glucose uptake and lactate production rates compared to those cultured under healthy plasma-like conditions (Mouse Plasma-like Medium, pH 7.4, 5% O2), despite lower initial glucose concentrations in TMEM (2.3 mM vs. 4.4 mM in Mouse Plasma-like Medium). Moreover, cell proliferation rates were lower in TMEM, suggesting an overall reduction in ATP consumption fueling cell division. Seahorse Extracellular Flux Analysis is being performed to determine effects on cellular bioenergetics. Metabolomic amino acid uptake using LCMS is also being performed to further assess the effect of TMEM on nutrient utilization. Using RNA-sequencing, we showed that culture in TMEM increased expression of genes associated with angiogenesis, amino acids biosynthesis, cell migration, and endothelial-to-mesenchymal transition. Migration assays confirmed increased motility in KPCY cells cultured in TMEM. Taken together, these results demonstrate that growth in a medium designed to simulate the TME alters KPCY cell biology in ways that are relevant to their pathology, which highlights the significance of maintaining TME-like conditions in culture. This research was funded by a Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant from JAS and a NSERC Canadian Graduate Student (Masters) Grant awarded to GLG. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,001 | 0,001 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,001 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,001 | 0,001 |
| Science ouverte | 0,000 | 0,001 |
| Intégrité de la recherche | 0,001 | 0,001 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,002 | 0,001 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule source (Gemma direct ou Codex distillé), pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».