Abstract
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.
Highlights
The urothelium is a transitional epithelium, lining on the inside of the bladder, ureters, and urethra, as well as the renal pelvis
Ancillary tests have been used for urine cytology, but only a few are currently approved by both Food and Drugs Administration (FDA) and European Association of Urology guidelines to be used in laboratory, as UroVysion Bladder Cancer Kit (Abbott Molecular, Des Plaines, IL, USA) [7,12]
Our preliminary results showed that this novel CytoMatrix protocol enables the rapid characterization of urothelial cancer cells after the urine specimens preparation, with the advantage of using ancillary tests to support cytological diagnosis and the advantage of storing cellular material in a formalinfixed paraffin embedded (FFPE) block for further analysis
Summary
The urothelium is a transitional epithelium, lining on the inside of the bladder, ureters, and urethra, as well as the renal pelvis. Few studies have demonstrated that urine cytology has a disappointingly low sensitivity for bladder cancer detection and the improvement of laboratory tests are needed [10,11] For these reasons, ancillary tests have been used for urine cytology, but only a few are currently approved by both Food and Drugs Administration (FDA) and European Association of Urology guidelines to be used in laboratory, as UroVysion Bladder Cancer Kit (Abbott Molecular, Des Plaines, IL, USA) [7,12]. Our preliminary results showed that this novel CytoMatrix protocol enables the rapid characterization of urothelial cancer cells after the urine specimens preparation, with the advantage of using ancillary tests to support cytological diagnosis and the advantage of storing cellular material in a formalinfixed paraffin embedded (FFPE) block for further analysis. Square Cover Slip (18 × 18 mm) (Thermo Fisher Scientific, Cat. no.: 18 × 18-1.5)
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