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Background: There are few studies on stem cell isolation in wild animals that provide isolation and culture protocols of these cells in vitro. Among the wild species studied, we present the collared peccary (Tayassu tajacu) as a model with potential to obtain and use MSC in preclinical studies. These animals are phylogenetically close to the domestic pig, popularly known as peccaries and found naturally in South America, Central America and the South of the United States. The aim of the present study was to establish a protocol for the isolation, in vitro cell expansion, differentiation and assessment of the stromal MSC growth curve before and after thawing.Materials, Methods & Results: Mesenchymal stem cells (MSC) from collared peccary bone marrow (Tayassu tajacu) were isolated and expanded by centrifuge in Ficoll® solution and cultured in DMEM® High Glucose medium. The culture was assessed by assays of colony forming units CFU-F and growth curve by saturation (GCS). Cultures in the third passage, with 70% confluence, were replicated at 105 cells/mL concentration in the culture media to induce osteogenic cell differentiation and adipogenic cell differentiation, respectively. The MSC were frozen in nitrogen for 40 days, thawed and re-assessed for cell viability and GCS.Discussion: The bone marrow collected presented high mononuclear cellularity, with a mean variability of 94.5% and 60.83 ± 4.27 UFC were identified in the samples and cells with fibroblast-like-cell morphology were observed. When they were expanded, the mean cell viability was 95%, the mean cell concentration obtained was 233.31 ± 20.04 cells per 25cm2 bottle and the culture reached the growth plateau in GCS between the 13th and 16th day. The osteoblastic cell differentiation assay showed after 18 days, morphology similar to osteoblasts, with irregular cytoplasm limits, cell prolongation formation and flattened appearance. After staining with Alizarin Red, the nucleus presented a wine red coloring and the cytoplasm, more basophilic and well-defined, with calcium deposits inside the cells. The cultures submitted to adipogenic differentiation were large, hexagonal, irregular and presented birrefringent cytoplasm granules after the third week of culture. When stained with Oil Red it was observed that the cytoplasm granules were scattered small fat vacuoles and stained maroon. The viability after thawing was 78% and the mean cell concentration obtained in GCS was 199.71 ± 14.72 cells per 25 cm2 bottle. The curves reached the saturation plateau early, on the eighth day of observation. From then onwards the cultures entered became exhausted and the cell concentration of the samples decreased progressively until minimum values. These results showed the presence of a well-defined MSC population in the collared peccary bone marrow with a high rate of replication in vitro and potential for differentiation confirmed by the adipogenic and osteogenic lines. The cryopreservation technique adopted presented satisfactory results, but indicated a significant cell stress after thawing that justifies investigation of the apoptosis rates induced post thawing in the species. Furthermore, the bone marrow collection did not harm the animals and the facility of stromal MSC isolation and culture qualifies the collared peccary as a viable alternative model to obtain MSC and for studies in the area of cell therapy.

The study aimed to isolate, expand, differentiate and characterize progenitor cells existent in the dental pulp of agouti. The material was washed with PBS solution and dissociated mechanically with the aid of a scalpel blade on plates containing culture medium D-MEM/F-12, and incubated at 5% CO2-37⁰C. The growth curve, CFU assay, osteogenic/adipogenic differentiation and characterization were obtained from the isolation. The cells began to be released from the explant tissue around the 7th day of culture. By day 22 of culture, cells reached 80% confluence. At the UFC test, 81 colonies were counted with 12 days of cultivation. The growth curves before and after freezing showed a regular growth with intense proliferation and clonogenic potential. The cell differentiation showed formation of osteoblasts and fat in culture, starting at 15 days of culture in a specific medium. Flow cytometry (FACs) was as follows: CD34 (positive), CD14 (negative), CD45 (negative), CD73 (positive), CD79 (negative), CD90 (positive), CD105 (positive), demonstrating high specificity and commitment of isolated cells with mesenchymal stem cells strains. These results suggest the existence of a cell population of stem cells with mesenchymal features from the isolated tissue in the explants of agouti dental pulp, a potential model for study of stem cell strains obtained from the pulp tissue.

; CARVALHO, M. A. M. de.; PESSOA, G. T.; FERRAZ, M. S.; PINTO, L. S. S. Citologia e histopatologia de pacientes assistidas em um centro de saúde da mulher. Arq. Ciênc. Saúde Unipar, Umuarama, v. 18 n. 1, p. 3-7, jan./ abr. 2014 RESUMO: O carcinoma de colo do útero é o segundo mais frequente em mulheres no Brasil, tendo acometido mais de 18.000 mulheres no ano de 2008. O presente estudo objetivou analisar e correlacionar os resultados obtidos nos laudos citológicos e histopatológicos, conforme a faixa etária das pacientes. Para tanto, metodologicamente foi realizado um estudo observacional transversal retrospectivo por meio de 106 prontuários de pacientes que apresentavam laudos citopatológicos e histopatológicos com atipias cervicais atendidas no período de janeiro de 2007 a dezembro de 2008 no Centro de Saúde da Mulher em Piripiri, Piauí, Brasil. Foram obtidos os seguintes resultados: quanto ao exame citológico os resultados mais frequentes foram os de ASCH e ASC/AG, correspondendo a respectivamente, 39,6% e 30%. Houve maior número de pacientes com laudo de ASC/AG e ASCH no intervalo de 25 a 34 anos, correspondendo a respectivamente, 13,2% e 17,9% destas lesões. Na histopatologia, 44 casos (41,5%) foram considerados LSIL, 53 casos (50%) foram diagnosticados com HSIL e 7 casos (6,6%) foram considerados como neoplasia maligna invasiva. Sendo assim, este estudo confirma a acuidade diagnóstica dos exames citológicos e histopatológicos, tendo em vista o grande número de lesões observadas, especialmente em pacientes jovens, destaca-se a importância de orientar práticas sexuais seguras, melhor controle na frequência de rastreamento e seguimento clínico destas pacientes. PALAVRAS-CHAVE: Citologia; Histopatologia; Prevenção do câncer do colo do útero. CYTOLOGY AND HISTOPATHOLOGY OF PATIENTS ASSISTED IN A WOMAN HEALTH CENTERABSTRACT: Cervical carcinoma is the second most common carcinomas among women in Brazil, having involved over 18,000 women in 2008. Analyze and correlate the most frequent cervical lesions observed in cytological and histological examinations. A cross -sectional observational study was retrospectively performed with 106 patients from a Woman Health Center in Piripiri, Piaui, Brazil, from January 2007 to December 2008, who presented atypical cytopathologic and histopathologic cervical findings. The cytologic results were more frequent in ASC and ASCH/AG, representing 39.6% and 30%, respectively. There was a greater number of patients with report of ASC/ AG and ASCH in the range of 25 to 34, representing respectively 13.2% and 17.9% of these lesions. Histopathologically, 44 cases (41.5%) were considered LSIL, 53 cases (50%) were diagnosed as HSIL and 7 cases (6.6%) were considered invasive malignancy. This study confirms the diagnostic accuracy of Pap smears, colposcopy and histopathology exams. Given the large number of lesions, especially in young patients, it highlights the importance of guidelines for safe sex practices, better control in the screening frequency and follow-up for these patients....

Stem cells are present in the adult tissues of most diverse species. Bone marrow is recognized to be the most exploited site to obtain stem cells and cell progenitors. The objective of the present study was to characterize hematopoietic progenitor (HP) morphology and analyze the performance of adherent cell progenitors (ACPs) cultivated in vitro from black-rumped agouti bone marrow (Dasyprocta prymnolopha). Bone marrow aspirates were obtained from tibia crest and used to prepare histological slides and identify cell morphology. Cells were also scattered on culture plates for later isolation, expansion, and quantification. Smears obtained from bone marrow demonstrated HPs at different stages of maturity. In culture, these cells showed fibroblastoid morphology and a strong tendency to form colonies, demonstrated by the presence of cell aggregates, cytoplasmic elongations lying side by side. An 80% cell confluence was observed at 18 days in culture and progressive reduction in the percentage of nonadherent mononuclear cells. After eight passes, a mean cell viability of 96.07% was observed, from a pool of 1.6 × 10(7) cells (ACP). Thirteen 25-cm(2) culture bottles were trypsinized, resuspended in freezing medium, stored in 14 criotubes at a concentration of 1 × 10(6) cells per milliliter, and placed in liquid nitrogen at -196°C. Agouti bone marrow demonstrated high plasticity, moreover different HP lines, and a population of adherent cells demonstrated morphology similar to mesenchymal stem cells in culture.