Cellusim: Un simulador 3D en entorno videojuego para la docencia del laboratorio de cultivos celulares

[EN] Cell cultures allow the maintenance of living cells outside the body. This technique is fundamental to study the biological, biochemical and physiological properties of cells and can be used as an experimental model in vitro in the field of biomedical research. The teaching in the laboratory of this technique presents numerous difficulties at a practical, logistical and economic level. On the other hand, the learning of cell culture protocols is an important part of the student's training in Cell Biology. Based on the routine practices carried out within a cell culture laboratory, we have developed the virtual 3D simulator of a cell culture laboratory "Cellusim". In "Cellusim" you can perform basic tasks of the cell culture of an established cell line of your own (Mel-RC08, Gil-Benso et al., 2012). To make Cellusim more attractive and intuitive to students, it has been developed in the Unity graphic environment, commonly used for the development of video games. The system includes the simulator in Unity, a mysql server with users, passwords and logs and a web of help and explanatory videos. In the simulator the student can virtually execute the essential tasks that are performed in a cell culture laboratory, such as thawing cells, seeding the cells, preparing and changing culture media and making cell subcultures. Cellusim is a training tool that allows users to discover the basics of cell culture techniques in a simple and fast way and without the economic costs or time consumption derived from doing the same work in a real laboratory. In order to evaluate the students' learning, Cellusim can be executed in training mode and in evaluation mode, allowing the student to perform all the processes as many times as necessary to become familiar with the protocols and when be able to execute them in evaluation mode. In evaluation mode, Cellusim records student errors in order to score the acquired training. In this communication we present the Cellusim project and the results of the first experiences with volunteer students of Master's Degrees in the School of Medicine.[ES] Los cultivos celulares permiten el mantenimiento de células vivas fuera del organismo. Esta técnica es fundamental para estudiar las propiedades biológicas, bioquímicas y fisiológicas de las células y puede ser utilizada como modelo experimental in vitro en el ámbito de la investigación biomédica. La enseñanza en el laboratorio de esta técnica presenta numerosas dificultades a nivel práctico, logístico y económico. Por otra parte, el aprendizaje de los protocolos del cultivo celular resultan una parte importante de la formación del alumno de Biología Celular. Basándonos en las prácticas rutinarias llevadas a cabo dentro de un laboratorio de cultivos celulares, hemos desarrollado el simulador virtual 3D de un laboratorio de cultivos celulares “Cellusim”. En “Cellusim” se pueden realizar tareas básicas del cultivo celular de una línea celular establecida propia (Mel‐RC08, Gil‐Benso y cols., 2012). Para hacer Cellusim más atractivo e intuitivo a los alumnos, ha sido desarrollado en el entorno gráfico Unity, utilizado habitualmente para el desarrollo de videojuegos. El sistema incluye el simulador en Unity, un servidor mysql con usuarios, contraseñas y registros y una web de ayuda y videos explicativos. En el simulador el alumno puede ejecutar de modo virtual las tareas esenciales que se realizan en un laboratorio de cultivos celulares, tales como la descongelación de células, el sembrado de las células, la preparación y el cambio de medios de cultivo y el subcultivo celular o técnica de doblaje. Cellusim es una herramienta formativa que permite a los usuarios descubrir los fundamentos de las técnicas básicas de cultivo celular de una manera sencilla, rápida y sin los costes económicos ni el consumo de tiempo derivados de realizar el mismo trabajo en un laboratorio real. Para poder evaluar el aprendizaje de los alumnos, Cellusim puede ser ejecutado en modo entrenamiento y en modo evaluación, permitiendo que el alumno realice todos los procesos las veces que sea necesario para familiarizarse con los protocolos y que cuando esté en condiciones pueda ejecutarlas en modo evaluación. En modo evaluación, Cellusim registra los errores del alumno para poder puntuar la formación adquirida. En esta comunicación presentamos el proyecto Cellusim y los resultados de las primeras experiencias con alumnos voluntarios de Master de la Facultad de Medicina.Monleón, D.; Megías, J.; San Miguel, T.; Borrás, C.; Gil Benso, R.; López Ginés, C. (2018). Cellusim: Un simulador 3D en entorno videojuego para la docencia del laboratorio de cultivos celulares. En IN-RED 2018. IV Congreso Nacional de Innovación Educativa y Docencia en Red. Editorial Universitat Politècnica de València. 176-183. https://doi.org/10.4995/INRED2018.2018.8714OCS17618

Molecular characterization and clinical impact of TMPRSS2-ERG rearrangement on prostate cancer: comparison between FISH and RT-PCR

Prostate cancer (PCa) is a very heterogeneous disease, and there are constraints in its current diagnosis. Serum PSA levels, digital rectal examination (DRE), and histopathologic analysis often drive to overdiagnosis and overtreatment. Since 2005, the presence of the genetic rearrangement between transmembrane-serine protease gene (TMPRSS2) and the erythroblast transformation-specific (ETS)member ERG (v-ets erythroblastosis virus E26 oncogene homolog avian) has been demonstrated in almost half of PCa cases. Both FISH and RT-PCR are useful tools for detecting these rearrangements, but very few comparatives between both techniques have been published. In this study, we included FFPE tumors from 294 PCa patients treated with radical prostatectomy with more than 5 years of followup.We constructed a total of 20 tissue microarrays in order to perform break-apart and tricolor probe FISH approaches that were compared with RT-PCR, showing a concordance of 80.6% ( P < 0.001). The presence of TMPRSS2-ERG rearrangement was observed in 56.6% of cases. No association between TMPRSS2-ERG status and clinicopathological parameters nor biochemical progression and clinical progression free survival was found. In conclusion, this study demonstrates that both FISH and RT-PCR are useful tools in the assessment of the TMPRSS2-ERG fusion gene status in PCa patients and that this genetic feature per se lacks prognostic value.This study has been funded by the Grants FIS PI06/01619 and PI10/01206 from the Instituto de Salud Carlos III, Madrid, ACOMP 12/029 from the Generalitat Valenciana, Valencia, and Astra Zeneca, Spain

Intracellular coexpression of CXC- and CC– chemokine receptors and their ligands in human melanoma cell lines and dynamic variations after xenotransplantation

BackgroundChemokines have been implicated in tumor progression and metastasis. In melanoma, chemokine receptors have been implicated in organ selective metastasis by regulating processes such as chemoattraction, adhesion and survival.MethodsIn this study we have analyzed, using flow cytometry, the systems formed by the chemokine receptors CXCR3, CXCR4, CXCR7, CCR7 and CCR10 and their ligands in thirteen human melanoma cell lines (five established from primary tumors and eight established from metastasis from different tissues). WM-115 and WM-266.4 melanoma cell lines (obtained from a primary and a metastatic melanoma respectively) were xenografted in nude mice and the tumors and cell lines derived from them were also analyzed.ResultsOur results show that the melanoma cell lines do not express or express in a low degree the chemokine receptors on their cell surface. However, melanoma cell lines show intracellular expression of all the aforementioned receptors and most of their respective ligands. When analyzing the xenografts and the cell lines obtained from them we found variations in the intracellular expression of chemokines and chemokine receptors that differed between the primary and metastatic cell lines. However, as well as in the original cell lines, minute or no expression of the chemokine receptors was observed at the cell surface.ConclusionsCoexpression of chemokine receptors and their ligands was found in human melanoma cell lines. However, this expression is intracellular and receptors are not found at the cell membrane nor chemokines are secreted to the cell medium. The levels of expressed chemokine receptors and their ligands show dynamic variations after xenotransplantation that differ depending on the origin of the cell line (from primary tumor or from metastasis)

Molecular Progression in Unusual Recurrent Non-Pediatric Intracranial Clear Cell Meningioma

We report a case of a recurrent clear cell meningioma (CCM) in the frontal lobe of the brain of a 67-year-old man. The patient developed three recurrences: at 3, 10, and 12 years after his initial surgery. Histopathology observations revealed a grade 2 CCM with positivity for vimentin and epithelial membrane antigen. Expression of E-cadherin was positive only in the primary tumour and in the first available recurrence. Fluorescence in situ hybridization analyses demonstrated 1p and 14q deletions within the last recurrence. Multiplex ligation-dependent probe amplification studies revealed a heterozygous partial NF2 gene deletion, which progressed to total loss in the last recurrence. The last recurrence showed homozygous deletions in CDKN2A and CDKN2B. The RASSF1 gene was hypermethylated during tumour evolution. In this report, we show the genetic alterations of a primary ccm and its recurrences to elucidate their relationships with the changes involved in the progression of this rare neoplasm

Correlation between EGFR amplification and the expression of microRNA-200c in primary glioblastoma multiforme.

Extensive infiltration of the surrounding healthy brain tissue is a critical feature in glioblastoma. Several miRNAs have been related to gliomagenesis, some of them related with the EGFR pathway. We have evaluated whole-genome miRNA expression profiling associated with different EGFR amplification patterns, studied by fluorescence in situ hybridization in tissue microarrays, of 30 cases of primary glioblastoma multiforme, whose clinicopathological and immunohistochemical features have also been analyzed. MicroRNA-200c showed a very significant difference between tumors having or not EGFR amplification. This microRNA plays an important role in epithelial-mesenchymal transition, but its implication in the behavior of glioblastoma is largely unknown. With respect to EGFR status our cases were categorized into three groups: high level EGFR amplification, low level EGFR amplification, and no EGFR amplification. Our results showed that microRNA-200c and E-cadherin expression are down-regulated, while ZEB1 is up-regulated, when tumors showed a high level of EGFR amplification. Conversely, ZEB1 mRNA expression levels were significantly lower in the group of tumors without EGFR amplification. Tumors with a low level of EGFR amplification showed ZEB1 expression levels comparable to those detected in the group with a high level of amplification. In this study we provide what is to our knowledge the first report of association between microRNA-200c and EGFR amplification in glioblastomas

Introducción a las alteraciones estructurales equilibradas de los cromosomas

Proyecto orientado al alumnado de Medicina. Se persigue, por un lado facilitar material complementario multimedia para preparar y entender mejor los supuestos prácticas que se plantean en la asignatura de Embriología, y por otro lado, consolidar los conocimientos que se van a desarrollar a lo largo de la materia. Música: Firefly by Techsmith library. Producción: Unidad de Biología. Con apoyo del SFPIE UV-SFPIE_RMD17-72497

Segregación cromosómica de la translocación robertsoniana

Proyecto orientado al alumnado de Medicina. Se persigue, por un lado facilitar material complementario multimedia para preparar y entender mejor los supuestos prácticos que se plantean en la asignatura de Embriología, y por otro lado, consolidar los conocimientos que se van a desarrollar a lo largo de la materia. Música: Firefly by Techsmith library. Producción: Unidad de Biología. Con apoyo del SFPIE UV-SFPIE_RMD17-72497

Gene Expression Profiles of Metabolic Aggressiveness and Tumor Recurrence in Benign Meningioma

<div><p>Around 20% of meningiomas histologically benign may be clinically aggressive and recur. This strongly affects management of meningioma patients. There is a need to evaluate the potential aggressiveness of an individual meningioma. Additional criteria for better classification of meningiomas will improve clinical decisions as well as patient follow up strategy after surgery. The aim of this study was to determine the relationship between gene expression profiles and new metabolic subgroups of benign meningioma with potential clinical relevance. Forty benign and fourteen atypical meningioma tissue samples were included in the study. We obtained metabolic profiles by NMR and recurrence after surgery information for all of them. We measured gene expression by oligonucleotide microarray measurements on 19 of them. To our knowledge, this is the first time that distinct gene expression profiles are reported for benign meningioma molecular subgroups with clinical correlation. Our results show that metabolic aggressiveness in otherwise histological benign meningioma proceeds mostly through alterations in the expression of genes involved in the regulation of transcription, mainly the LMO3 gene. Genes involved in tumor metabolism, like IGF1R, are also differentially expressed in those meningioma subgroups with higher rates of membrane turnover, higher energy demand and increased resistance to apoptosis. These new subgroups of benign meningiomas exhibit different rates of recurrence. This work shows that benign meningioma with metabolic aggressiveness constitute a subgroup of potentially recurrent tumors in which alterations in genes regulating critical features of aggressiveness, like increased angiogenesis or cell invasion, are still no predominant. The determination of these gene expression biosignatures may allow the early detection of clinically aggressive tumors.</p></div