Preclinical studies based on a "simulation design", were performed with cultured melanoma cells prelabeled with 51Cr, added to normal blood and subjected to separation and recognition steps. Mononuclear cells (MNC) were isolated on ficollhypaque gradient, and melanoma cells were separated from lymphocytes using anti-CD45 immunomagnetic beads. Malignant cells were then recognized by measuring telomerase activity (TRAP and TRAP-ELISA assays). It was found that: (a) recovery of prelabeled cells present in MNC did not exceed 75%; (b) further recovery of prelabeled cells after separation from lymphocytes did not exceed 68%. Therefore, the overall recovery of prelabeled cells did not exceed 48%; (c) the entire procedure was able to reliably detect as few as 30 malignant cells added to normal blood, providing a telomerase signal significantly higher than that found in absence of melanoma cells. These results furnish the technical bases for developing a tumor detection assay in the blood of melanoma patients.

Bonmassar, L., Massara, M.C., Cottarelli, A., Aquino, A., Formica, V., Prete, S.P., et al. (2004). Preclinical studies on detection of circulating melanoma cells in patients: Telomerase as a recognition marker of malignancy. JOURNAL OF CHEMOTHERAPY, 16(5), 479-486.

Preclinical studies on detection of circulating melanoma cells in patients: Telomerase as a recognition marker of malignancy

AQUINO, ANGELO;PRETE, SALVATORE;FARAONI, ISABELLA;
2004

Abstract

Preclinical studies based on a "simulation design", were performed with cultured melanoma cells prelabeled with 51Cr, added to normal blood and subjected to separation and recognition steps. Mononuclear cells (MNC) were isolated on ficollhypaque gradient, and melanoma cells were separated from lymphocytes using anti-CD45 immunomagnetic beads. Malignant cells were then recognized by measuring telomerase activity (TRAP and TRAP-ELISA assays). It was found that: (a) recovery of prelabeled cells present in MNC did not exceed 75%; (b) further recovery of prelabeled cells after separation from lymphocytes did not exceed 68%. Therefore, the overall recovery of prelabeled cells did not exceed 48%; (c) the entire procedure was able to reliably detect as few as 30 malignant cells added to normal blood, providing a telomerase signal significantly higher than that found in absence of melanoma cells. These results furnish the technical bases for developing a tumor detection assay in the blood of melanoma patients.
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore BIO/14
Settore MED/06 - Oncologia Medica
eng
Con Impact Factor ISI
CD45; Circulating cells; Immunomagnetic beads; Melanoma; Telomerase; TRAP Assay
Bonmassar, L., Massara, M.C., Cottarelli, A., Aquino, A., Formica, V., Prete, S.P., et al. (2004). Preclinical studies on detection of circulating melanoma cells in patients: Telomerase as a recognition marker of malignancy. JOURNAL OF CHEMOTHERAPY, 16(5), 479-486.
Bonmassar, L; Massara, M; Cottarelli, A; Aquino, A; Formica, V; Prete, S; Lacal, P; Marchetti, P; Concolino, F; Faraoni, I; D'Atri, S
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2108/40649
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