Improvement of the Collection, Maintenance, and Analysis of Neoplastic Cells from Urine Specimens with the Use of CytoMatrix

Urine cytology is a non-invasive test used in combination with cystoscopy for screening and follow-up of urothelial carcinoma (UC). Although cytology can be used to efficiently identify high-grade UC, it has a lower accuracy for the diagnosis of low-grade UC or patients with presence of atypical uro...

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Main Authors: Simone Minasi, Daniela Bosco, Bernardo Moretti, Felice Giangaspero, Antonio Santoro, Francesca Romana Buttarelli
Format: Article
Language:English
Published: MDPI AG 2021-09-01
Series:Methods and Protocols
Subjects:
Online Access:https://www.mdpi.com/2409-9279/4/3/65
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author Simone Minasi
Daniela Bosco
Bernardo Moretti
Felice Giangaspero
Antonio Santoro
Francesca Romana Buttarelli
author_facet Simone Minasi
Daniela Bosco
Bernardo Moretti
Felice Giangaspero
Antonio Santoro
Francesca Romana Buttarelli
author_sort Simone Minasi
collection DOAJ
description Urine cytology is a non-invasive test used in combination with cystoscopy for screening and follow-up of urothelial carcinoma (UC). Although cytology can be used to efficiently identify high-grade UC, it has a lower accuracy for the diagnosis of low-grade UC or patients with presence of atypical urothelial cells (AUC). For these reasons, ancillary tests have been added to urine cytology in order to improve the accuracy. However, the poor abundance of neoplastic cells in most samples and the absence of a “tissue-like” structure remains a major challenge. We used a novel synthetic support called CytoMatrix which has the property of capturing and storing cells and micro-macro aggregates within its three-dimensional structure. The urine specimens were obtained from 12 patients: 6 with suspected urothelial neoplasia (low- and high-grade) and 6 with AUC or non-neoplastic samples. The first step is the urine samples preparation, through several centrifugation passages; the second step consists in absorbing cells on the CytoMatrix, and in the subsequent formalin fixation, standard processing and paraffin embedding to prepare FFPE-CytoMatrix block. In the final step, sections are consecutively cut, stained with hematoxylin-eosin (H&E), and analyzed via UroVysion FISH and immunohistochemistry (IHC). Using our simple and reliable protocol, we can improve the quality of urine specimens, allowing a better collection, maintenance, and analysis of cells, with the advantage of using ancillary tests to support cytological diagnosis and the advantage of storing cellular material in a FFPE-CytoMatrix block.
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spelling doaj.art-03051892fa7d4eaaa28be11b693b24502023-11-22T14:27:53ZengMDPI AGMethods and Protocols2409-92792021-09-01436510.3390/mps4030065Improvement of the Collection, Maintenance, and Analysis of Neoplastic Cells from Urine Specimens with the Use of CytoMatrixSimone Minasi0Daniela Bosco1Bernardo Moretti2Felice Giangaspero3Antonio Santoro4Francesca Romana Buttarelli5Department of Radiological, Oncological and Anatomo-Pathological Sciences, Sapienza University of Rome, 00161 Roma, ItalyDepartment of Radiological, Oncological and Anatomo-Pathological Sciences, Sapienza University of Rome, 00161 Roma, ItalyDepartment of Radiological, Oncological and Anatomo-Pathological Sciences, Sapienza University of Rome, 00161 Roma, ItalyDepartment of Radiological, Oncological and Anatomo-Pathological Sciences, Sapienza University of Rome, 00161 Roma, ItalyUCS Diagnostic S.r.l., 00067 Rome, ItalyDepartment of Radiological, Oncological and Anatomo-Pathological Sciences, Sapienza University of Rome, 00161 Roma, ItalyUrine cytology is a non-invasive test used in combination with cystoscopy for screening and follow-up of urothelial carcinoma (UC). Although cytology can be used to efficiently identify high-grade UC, it has a lower accuracy for the diagnosis of low-grade UC or patients with presence of atypical urothelial cells (AUC). For these reasons, ancillary tests have been added to urine cytology in order to improve the accuracy. However, the poor abundance of neoplastic cells in most samples and the absence of a “tissue-like” structure remains a major challenge. We used a novel synthetic support called CytoMatrix which has the property of capturing and storing cells and micro-macro aggregates within its three-dimensional structure. The urine specimens were obtained from 12 patients: 6 with suspected urothelial neoplasia (low- and high-grade) and 6 with AUC or non-neoplastic samples. The first step is the urine samples preparation, through several centrifugation passages; the second step consists in absorbing cells on the CytoMatrix, and in the subsequent formalin fixation, standard processing and paraffin embedding to prepare FFPE-CytoMatrix block. In the final step, sections are consecutively cut, stained with hematoxylin-eosin (H&E), and analyzed via UroVysion FISH and immunohistochemistry (IHC). Using our simple and reliable protocol, we can improve the quality of urine specimens, allowing a better collection, maintenance, and analysis of cells, with the advantage of using ancillary tests to support cytological diagnosis and the advantage of storing cellular material in a FFPE-CytoMatrix block.https://www.mdpi.com/2409-9279/4/3/65urothelial carcinomabladder cancerurineUroVysionCytoMatrix
spellingShingle Simone Minasi
Daniela Bosco
Bernardo Moretti
Felice Giangaspero
Antonio Santoro
Francesca Romana Buttarelli
Improvement of the Collection, Maintenance, and Analysis of Neoplastic Cells from Urine Specimens with the Use of CytoMatrix
Methods and Protocols
urothelial carcinoma
bladder cancer
urine
UroVysion
CytoMatrix
title Improvement of the Collection, Maintenance, and Analysis of Neoplastic Cells from Urine Specimens with the Use of CytoMatrix
title_full Improvement of the Collection, Maintenance, and Analysis of Neoplastic Cells from Urine Specimens with the Use of CytoMatrix
title_fullStr Improvement of the Collection, Maintenance, and Analysis of Neoplastic Cells from Urine Specimens with the Use of CytoMatrix
title_full_unstemmed Improvement of the Collection, Maintenance, and Analysis of Neoplastic Cells from Urine Specimens with the Use of CytoMatrix
title_short Improvement of the Collection, Maintenance, and Analysis of Neoplastic Cells from Urine Specimens with the Use of CytoMatrix
title_sort improvement of the collection maintenance and analysis of neoplastic cells from urine specimens with the use of cytomatrix
topic urothelial carcinoma
bladder cancer
urine
UroVysion
CytoMatrix
url https://www.mdpi.com/2409-9279/4/3/65
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AT bernardomoretti improvementofthecollectionmaintenanceandanalysisofneoplasticcellsfromurinespecimenswiththeuseofcytomatrix
AT felicegiangaspero improvementofthecollectionmaintenanceandanalysisofneoplasticcellsfromurinespecimenswiththeuseofcytomatrix
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