Abstract 2912: Size based enrichment and sorting of Ov90 cancer cells and clusters with a new multistage filtration cartridge reveals distinct phenotypes
Notice bibliographique
Résumé
Abstract Background: Circulating tumor cells (CTCs) are released in blood from the primary tumor, but although very heterogeneous both in size and marker expression, are very rare and provide information not available from the primary tumor. The identification of CTC cells and clusters could advance our understanding of metastasis and help personalize therapy.1,2 Notably, CTC clusters were shown to have a higher metastatic potential than single cells3 but the process remains poorly understood. We present a multistage filtration system with pore sizes from 20 down to 8 μm for the size-selective enrichment of Ov90 ovarian cancer cells and clusters from blood. Each captured cell population was released, cultured and characterized independently. Methods: We developed a 3D printed multistage filtration cartridge and polymer filters with 20, 15, 12, 10 and 8 μm-diameter pores. Filters were stacked from 20 (top) to 8 μm (bottom) and used to enrich and sort Ov-90 cells spiked in 1:6 mL of blood:PBS. Captured cells were released by removing individual filters from the cartridge, reverse flowing OSE medium, and then cultured separately. Results: Ov-90 clusters were found mostly on the top filter (20 μm) and interestingly, few small clusters (3-4 cells) were found on the 8 and 10 μm filters, suggesting alignment of cluster cells as they pass through the pores.4 Cell and nucleus diameters were measured, and a general correlation was found between filter pore size and cell and nucleus size. Interestingly, nucleus size was found to be the single most significant parameter in determining passage of single cells and small clusters through pores. Following cell culture, two distinct phenotypes were observed: cell captured on small pore filters (8-12 μm) grew primarily in a monolayer. Cells captured on filters with larger pores (15-20 μm) first grew as monolayer, but rapidly formed cell aggregates that subsequently detached from the surface. Staining for E-Cadherin, a cell-cell adhesion protein, revealed a loss of expression of cells from filter with larger pores. Conclusion: We developed a new multistage filtration method and selectively enriched and sorted cells based primarily on their nucleus size. We identified two Ov-90 populations with different growth behaviors with low E-cadherin expression on the cells forming clusters, which is known to correlate with metastasis. The application of multistage filters may also reveal different CTC populations based on nucleus and cell size. References: (1) Baccelli, I. et al. A. Nat. Biotech. 2013, 31, 539-544. (2) Pecot, C. V. et al. Cancer discovery 2011, 1, 580-586. (3) Cheung, K. J. et al. Proc. Natl. Acad. Sci. U.S.A. 2016, 113, E854-E863. (4) Au, S. H. et al. Proc. Natl. Acad. Sci. U.S.A. 2016, 113, 4947-4952. Citation Format: Anne Meunier, Sara kheireddine, J. Alejandro Hernández-Castro, Teodor Veres, David Juncker. Size based enrichment and sorting of Ov90 cancer cells and clusters with a new multistage filtration cartridge reveals distinct phenotypes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2912. doi:10.1158/1538-7445.AM2017-2912
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 | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,001 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,001 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,002 | 0,002 |
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 ».