Open Journal of Veterinary Medicine, 2012, 2, 216-224 http://dx.doi.org/10.4236/ojvm.2012.24034 Published Online December 2012 (http://www.SciRP.org/journal/ojvm) Evaluation of Collection and Distribution of Samples for Histological, Stereological Analysis and Cell Culture of Canine Mast Cell Tumors * Samanta Rios Melo, Thaís Andrade Costa-Casagrande, Julia Maria Matera Department of Surgery, School of Veterinary Medicine, University of São Paulo, São Paulo, Brazil Email: samymelo@usp.br Received September 5, 2012; revised November 1, 2012; accepted November 12, 2012 ABSTRACT Objective: To collect high quality, representative tissue material from tumors and manage its distribution to different laboratories. Design: Prospective controlled study. Animals: Thirty-six dogs with mast cell tumors. Procedures: The samples were submitted for the following analyses: stereology; histopathology; cell culture; breakdown for cytogenetic analysis of chromosomes (based on the Boxer breed published genome); Cell lysis for Real Time PCR and quantifica- tion of gene expression of CX 43, 32 and E-cadherin in canine mast cells. Results: Cytogenetic chromosome analysis, 90.9% of the samples were considered to be of good quality. For gene expression quantification of CX 43, 32 and E-cadherin in canine mast cell tumors (MCT), 95.5% of samples were considered to be of good quality. Conclusions and Clinical Relevance: We seek to assess the importance of surgical collection and post-surgical tissue preparation on laboratory testing by collecting surgical material appropriately to allow accurate diagnosis and reliable clinical progno- sis and minimize errors caused by inadequate preparation of samples. Keywords: Mast Cell Tumor; Oncologic Surgery; Canine; Tumor Collection; Stereological Analysis 1. Introduction Mast cell tumors are common neoplasia in small animals. This type of neoplasm occurs in several species, includ- ing humans, but the major incidence is in canines and felines [1-3]. Mast cell tumors in the dog most commonly develop in the skin. These tumors usually have a variable appearance. In the predominant formation, the mast cell looks similar to a normal cell and is relatively easy to diagnose. Several approaches have been used to define prognosis for mast cell tumors and for other types of ca- nine and feline tumors. Clinical reports, surgical resec- tion [1,4,5] and histological analyses of the tissue, as well as assessment of kinetic parameters such as Ki-67 pro- tein [6,7], BrdU [7], PCNA [8], AgNOR [9]-Argyrophilic nucleolar organiser region, DNA ploidy[10], intratumoral vascular density [11], p53 [12,13], c-KIT [14,15], nu- clear morphometry [16], tumor profundity[17], and tu- mor localization [18]. The most common method used for determining the prognosis of a mast cell tumor is his- tological gradation [19]. The histological evaluation is also needed for the classification of the tumor and to ac- cess surgical margins. The diagnosis of mast cell tumors and other tumors depends on the pathologist’s access to the material col- lected from the surgeon. Most cancer diagnoses are de- termined using cytological (fine need aspiration) and histological (excisional biopsy) evaluations. Consequently, mistakes in the pathological evaluation can lead to inap- propriate treatment choices [20,21]. Several methods exist for studying mistakes in anatomopathological evalu- ation of the samples submitted for analysis. Sampling mistakes are most commonly related to the preparation of the samples (inappropriate storage of the samples, lack of anatomical references for the fragments, and insufficient samples from small tumors) [20-22]. The officially released percentage of error is known to vary significantly, ranging from 1% to 43% (averaging between 1% and 5%) in human medicine [22]. Failures are known to originate from the surgeon or his/her team upon transfer of the material for histopathology. In the literature, no studies in human or veterinary medicine have quantified the percentage of error in the histopa- thological analysis that results from inappropriate prepa- ration of the samples. Excisional biopsy is used to provide useful informa- tion and has diagnostic and prognostic roles in several types of human and canine tumors.. Some care must be * Supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP). Presented as an abstract at the 30th Annual Con- ference of the Veterinary Cancer Society, San Diego—USA, 2010. Copyright © 2012 SciRes. OJVM