Abstract 511: Unbiased enrichment of circulating tumor cells directly from whole blood
Notice bibliographique
Résumé
Abstract Enrichment of rare circulating tumor cells (CTC) in peripheral blood is required prior to most analytic procedures. The ideal enrichment method would be rapid (<30 minutes (min)), permit a high recovery of viable CTC, and would be independent of surface marker expression, since CTC in the peripheral blood may be undergoing epithelial-mesenchymal transition and may not express epithelial markers. We have developed a new immunomagnetic method (EasySep™ Direct) that fulfills these criteria and can be used directly with whole blood without any preparatory steps. The method was tested using whole blood samples seeded with the adenocarcinoma cell line CAMA to a final concentration of approximately 1%. The CAMA cells were then enriched using either EasySep™ Direct CTC Enrichment (immunomagnetic separation) or RosetteSep™ (immunodensity based control). For immunomagnetic enrichment, the samples were incubated with EasySep™ Direct Enrichment Cocktail for 5 min, then EasySep™ Direct RapidSpheres™ were added and the sample placed in a magnet for 10 min. Magnetically labeled cells were retained in the magnet and the unlabeled cells poured or pipetted off. Rapidspheres™ were added again and the magnetic incubation and pour-off steps repeated. To perform RosetteSep™ enrichment, the samples were incubated with either the CD45+ cell depletion cocktail or a more extensive hematopoietic cell depletion cocktail for 10 min. Samples were then pipetted onto Lymphoprep in SepMate™ tubes, centrifuged for 10 min at 1200 x g with the brake on, and poured off. The number of cells recovered with each procedure was counted and the depletion of hematopoietic cells (CD45+/EPCAM-), enrichment of CAMA cells (CD45-/EpCAM+), and depletion of red blood cells (RBCs, Glycophorin A+/CD45-) in each sample were evaluated by flow cytometry. EasySep™ Direct was evaluated on five different samples. Each sample was separated on five different EasySep™ magnet formats (5, 14, and 50 mL single tube magnets and 5 and 14 mL multi-tube magnets) with each condition in duplicate; the two RosetteSep™ cocktails were tested on three of the samples, each in duplicate. There was no significant difference in CD45 depletion, CAMA cell recovery, or residual RBC contamination between samples enriched using the different magnets (Least Squares Fit; Prob F>0.05) or pooled and compared to the pooled RosetteSep™ controls (Least Squares Fit; Prob F>0.05). Over all magnet separation conditions and samples, the mean log depletion of CD45+ cells was 2.9±0.4, the mean recovery of CAMA cells was 42±23%, and the residual RBC contamination was ∼9000 RBC/mL of start sample. Rare cells were enriched in 25 minutes using EasySep™ Direct and in less than 40 minutes using RosetteSep™ with SepMate™. CTC can be enriched directly from whole blood with either EasySep™ Direct or RosetteSep™. Since enriched cells are unlabeled there is nothing to interfere with subsequent further enrichment, culture, or evaluation. Citation Format: Carrie E. Peters, Danay Maestre-Battle, Steven M. Woodside, Terry E. Thomas, Allen C. Eaves. Unbiased enrichment of circulating tumor cells directly from whole blood. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 511.
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 | 0,001 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,001 | 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 | 0,000 | 0,001 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,003 | 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 ».