A Szentágothai János Kutatóközpont a PTE korszerű, nemzetközi tudományszervezési és menedzsment normák szerint kialakított új intézménye, amely az élettudományi, élettelen természettudományi, valamint környezettudományi oktatás...
Although cell culture and isolation of cells from tissues seems trivial at first, every cell culture is different. Depending on their origin, cells grow at a different rate, have different nutritional requirements, and have different periods of sustainability. Our cell lines include adherent cells and cells that can be maintained in suspension. Tumor cell lines are available to customers after they have been characterized (with knowledge of their mutations). Our cell lines include genetically modified cell lines that can be used in well-defined experiments. Our cell lines are of human or animal origin, we currently do not conduct experiments with plant cells.
The Cell and Tissue Culture Core Facility provides the opportunity to design, prepare (thaw, passage and handle specific cell lines), perform experiments and evaluate results based on personalized cell and tissue cultures using its comprehensive instruments and equipments. In addition to the experimental set-up based on two-dimensional cell and tissue cultures, users have the option to set up an experimental model based on complex cell and tissue cultures. Users are also provided with the ability to dissociate primary cells from animal or human tissues or to separate a particular cell population from body fluids.
The available cell cultures are cultured in all cases at 37 °C incubators with 5% CO2 and high humidity. The experiments are performed in a laminar flow cabinet under sterile conditions. In addition to the instruments highlighted below, we have the essential instruments and tools needed for cell and tissue culture techniques, such as:
Extracellular vesicles (EVs) are membrane-delimited particles released from all type of cells into the extracellular space. According to their size and biogenesis EVs can be differentiated into three subgroups such as exosomes, microvesicles and apoptotic bodies. These vesicles can be isolated from many body fluids (blood, urine, breast milk etc.) or from cell culture supernatant. Apoptotic bodies and microvesicles can be elutriated using high-speed centrifugation, but exosomes require PEG-based isolation method.
We are able to isolate exosomes from the following fluids:
In our laboratory, it is possible to characterize the isolated vesicles (based on size and quantity), which is performed with the Malvern NanoSight NS300 device using nanoparticle tracking analysis (NTA).
Esco biological safety cabinet provides appropriate environment for basic cell culture procedures, expreimental set-ups and ensures the required sterile condition.
The EuroClone s@femate 1.2 cyto safety cabinet allows safe handling of cytostatics in the Cell and Tissue Culture Core Facility, keeping both the operator and the sample as well as the environment protected thanks to the built-in 3 H14 HEPA filters. To use the cabinet, wearing protective clothing and following the rules for working with cytostatics is obligatory!
The Malvern Panalytical NanoSight NS300 Instrument provides an easy-to-use, reproducible platform for nanoparticle characterization, especially size characterization of extracellular vesicles. Particles in liquid suspension are loaded into a sample chamber. In the path of the beam the particles scatter the laser light which is easily collected by the microscope objective and is viewed with a digital camera. The camera captures a video of the particles moving under Brownian motion. The NS300 allows rapid analysis of the size distribution and concentration of all types of nanoparticles from 10 nm - 2 µm in diameter, depending on the instrument configuration and sample type. With the ability to be supplied with interchangeable laser modules and the introduction of a motorized filter wheel means different fluorescent labels can be analyzed.
Detectable particle size: 10nm - 2000nm
Lasers: 488nm (Blue), 532nm (Green)
Concentration: 109 particle / ml
NS300 can be applied in the development of different delivery system studies:
The nCounter SPRINT Profiler is a platform that allows you to measure nearly 800 targets simultaneously in 12 samples in just 7 hours. It is also able to analyze samples from relatively low concentrations: 1 - 50 ng for RNA, 5 - 300 ng for DNA, 250 ng - 2.5 μg for protein, which requires only a short sample preparation process.
The analysis of raw data can be done easily with the help of nSolver ™ Analysis Software.
More information about the instrument is available here.
Available panels and assays:
The QuantStudio™ 12K Flex Real-Time PCR system takes qPCR technology to a next level with maximum throughput and minimal resources. Using a simple workflow gene expression analysis or miRNA profiling can be conducted with reduced start-up and hands-on time. The results can be easily analyzed using ExpressionSuite Software, an easy-to-use data-analysis tool that utilizes the comparative Cτ (ΔΔCτ) method to rapidly and accurately quantify relative gene expression across a large number of genes and samples.
Fluorophore compatibility: Taqman and SYBR Green reagents
Interchangeable blocks and plate compatibility: TaqMan Array Card (384-well)
Fast 96-well plate
The gentleMACS™ Octo Dissociator is a benchtop instrument for semi-automated and standardized tissue dissociation or homogenization of up to eight samples. Single-cell suspensions or thorough homogenates from virtually any tissues are easily and reproducibly obtained using the unique C Tubes or M Tubes. These tubes allow sample preparation in a closed and sterile environment with minimized cross-contamination.
Optimized, pre-set gentleMACS Octo Dissociator Programs for easy dissociation of tissues into single-cell suspensions:
Greiner Bio-One's magnetic cell culturing technology uses biocompatible nanoparticles (NanoShuttle™-PL) in order to magnetize cells. After magnetization cells can form three-dimensional spheroid models in a relatively short period of time (24 hours). This high throughput technique is compatible with fluorescent microscopy, Western blot, qRT-PCR, flow cytometry, viability assays and chemiluminescence methods.
The advantages of magnetic cell culture include:
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The OctoMACS™ Separator contains a powerful permanent magnet that induces a high-gradient magnetic field to retain cells labeled with even small amounts of MACS MicroBeads. It allows the performance of up to eight simultaneous separations in combination with different MACS Columns:
Based on a cytospin method cells in cell suspension can be easily concentrated onto glass slides in order to perform different staining methods. The MPW-223c laboratory centrifuge is perfectly suitable to carry out such techniques.
Az EuroClone s@femate 1.5 eco biological safety cabinet provides appropriate environment for basic cell culture procedures, expreimental set-ups and ensures the required sterile condition.
The LAS 4000 is a camera system for producing digital images of chemiluminescent or fluorescent gel and membrane samples. The digital image allows the subsequent molar mass or quantitative analysis for e. g. Western blot.
Detection methods:
Exchangeable light sources and filters:
The QuantStudio 3D Digital PCR system is a chip-based technology which allows a simple and streamlined workflow with the ability to thermal cycle up to 24 chips at one time in less than 30 seconds to read each chip. The QuantStudio 3D Digital PCR System produces absolute quantification data, concentration measurements in copies/μL without a standard curve.
Applications:
Applied Biosystems™ QuantStudio™ 3D AnalysisSuite™ Cloud Software is used for the analysis of data derived from the QuantStudio 3D Digital PCR instrument. The software also enables analysis of QuantStudio 3D chips and accurate, highly precise measurements of genomic targets in copies per microliter. Moreover it allows low-fold detection comparison and ratio analysis of genomic targets, as well as copy number variation. Multiple chips can be combined and analyzed in a single project.
Voros-Horvath Barbara; Zivkovic Pavo; Banfai Krisztina; Bovari-Biri Judit; Pongracz Judit; Balint Gabor; Pal; Szechenyi Aleksandar: Preparation and Characterization of ACE2 Receptor Inhibitor-Loaded Chitosan Hydrogels for Nasal Formulation to Reduce the Risk of COVID-19 Viral Infection. ACS Omega 2022
Abdelwahab ElHusseiny M M; Judit Bovari-Biri; Gabor Smuk; Janos Fillinger; Donald McPhail; Vera P Krymskaya; Judit E Pongracz: Activated p53 in the anti-apoptotic milieu of tuberous sclerosis gene mutation induced diseases leads to cell death if thioredoxin reductase is inhibited. Apotosis 2021
Jaromi L.; Csongei V.; Vesel M.; Abdelwahab E.M.M.; Soltani A.; Torok Z.; Smuk G.; Sarosi V.; Pongracz J.E.: Kras and egfr mutations differentially alter abc drug transporter expression in cisplatin-resistant non-small cell lung cancer. International Journal of Molecular Sciences 2021
Abdelwahab Elhusseiny Mohamed Mahmoud; Bovari-Biri; Smuk Gabor; Harko Tunde; Fillinger Janos ; Moldvay Judit; Krymskaya Vera P; Pongracz Judit: Normalization of Enzyme Expression and Activity Regulating Vitamin A Metabolism Increases RAR-Beta Expression and Reduces Cellular Migration and Proliferation in Diseases Caused by Tuberous Sclerosis Gene Mutations. Frontiers in Oncology 2021
Csenki Zsolt; Garai Edina; Faisal Zelma; Csepregi Rita; Garai Kitti; Sipos Dóra Kánainé; Szabó István; Kőszegi Tamás; Czéh Árpád; Czömpöly Tamás; Kvell Krisztián; Poór Miklós: The individual and combined effects of ochratoxin A with citrinin and their metabolites (ochratoxin B, ochratoxin C, and dihydrocitrinone) on 2D/3D cell cultures, and zebrafish embryo models. Food and Chemical Toxicology 2021
Soltani A, Kajtar B, Abdelwahab EHMM, Steib A, Horvath Z, Mangel L, Jaromi L, Pongracz JE. Is an Immunosuppressive Microenvironment a Characteristic of Both Intra- and Extraparenchymal Central Nervous Tumors? Pathophysiology (2021) 28:34–49. doi:10.3390/pathophysiology28010004.
Garai Kitti; Adam Zoltan Robert Herczeg; Krisztina Banfai; Adam Gyebrovszki; Attila Gyenesei; Judit E Pongracz; Marta Wilhelm; Krisztian Kvell: Physical Activity as a Preventive Lifestyle Intervention Acts Through Specific Exosomal miRNA Species-Evidence From Human Short- and Long-Term Pilot Studies. Frontiers in Physiology 2021
Weich Alexander; Rogoll Dorothee; Gawlas Sophia; Mayer Lars; Weich Wolfgang; Pongracz Judit; Kudlich Theodor; Meining Alexander; Scheurlen Michael: Wnt/β-Catenin Signaling Regulates CXCR4 Expression and [68Ga] Pentixafor Internalization in Neuroendocrine Tumor Cells. Diagnostics 2021
Boda F, Banfai K, Garai K, Kovacs B, Almasi A, Scheffer D, Sinkler RL, Csonka R, Czompoly T, Kvell K. Effect of Bitis gabonica and Dendroaspis angusticeps snake venoms on apoptosis-related genes in human thymic epithelial cells. J Venom Anim Toxins Incl Trop Dis. 2020 Dec 14;26:e20200057. doi: 10.1590/1678-9199-JVATITD-2020-0057. PMID: 33402885; PMCID: PMC7745260.
Papp H, Zeghbib S, Földes F, Banfai K, Madai M, Kemenesi G, Urbán P, Kvell K, Jakab F. Crimean-Congo hemorrhagic fever virus infection triggers the upregulation of the Wnt signaling pathway inhibitor genes. Virus Genes. 2020 Aug;56(4):508-514. doi: 10.1007/s11262-020-01759-z. Epub 2020 Apr 25. PMID: 32335793.
Papp E, Steib A, Abdelwahab EM, Meggyes-Rapp J, Jakab L, Smuk G, Schlegl E, Moldvay J, Sárosi V, Pongracz JE. Feasibility study of in vitro drug sensitivity assay of advanced non-small cell lung adenocarcinomas. BMJ Open Respir Res. 2020 Jun;7(1):e000505. doi: 10.1136/bmjresp-2019-000505. PMID: 32527872; PMCID: PMC7292226.
Kiss E, Abdelwahab EHMM, Steib A, Papp E, Torok Z, Jakab L, Smuk G, Sarosi V, Pongracz JE. Cisplatin treatment induced interleukin 6 and 8 production alters lung adenocarcinoma cell migration in an oncogenic mutation dependent manner. Respir Res (2020) 21:120. doi:10.1186/s12931-020-01389-x
Abdelwahab, EMM; Rapp, J; Feller, D; Csongei, V; Pal, S; Bartis, D; Thickett, DR; Pongracz JE.: Wnt signaling regulates trans-differentiation of stem cell like type 2 alveolar epithelial cells to type 1 epithelial cells. Respiratory Research 20:1, 9 p. (2019)
Garai, K; Adam, Z; Herczeg, R; Katai, E; Nagy, T; Pal, S; Gyenesei, A; Pongracz, JE; Wilhelm, M; Kvell, K.: Artificial Neural Network Correlation and Biostatistics Evaluation of Physiological and Molecular Parameters in Healthy Young Individuals Performing Regular Exercise. Frontiers in Physiology 10 Paper: 1242, 11 p (2019)
Vas V, Háhner T, Kudlik G, Ernszt D, Kvell K, Kuti D, Kovács KJ, Tóvári J, Trexler M, Merő BL, Szeder B, Koprivanacz K, Buday L. Analysis of Tks4 Knockout Mice Suggests a Role for Tks4 in Adipose Tissue Homeostasis in the Context of Beigeing. Cells. 2019 Aug 5;8(8). pii: E831. doi: 10.3390/cells8080831.
Banfai K, Ernszt D, Pap A, Bai P, Garai K, Belharazem D, Pongracz JE, Kvell K. ‚Beige‘ Cross Talk between the Immune System and Metabolism. Front. Endocrinol.2019 May 24. doi: 10.3389/fendo.2019.00369.7
Banfai K, Garai K, Ernszt D, Pongracz JE, Kvell K. Transgenic Exosomes for Thymus Regeneration. Front Immunol. 2019 Apr 24;10:862. doi: 10.3389/fimmu.2019.00862.
Abdelwahab EMM, Pal S, Kvell K, Sarosi V, Bai P, Rue R, Krymskaya V, McPhail D, Porter A, Pongracz JE. Mitochondrial dysfunction is a key determinant of the rare disease lymphangioleiomyomatosis and provides a novel therapeutic target. Oncogene. 2019 Apr;38(16):3093-3101. doi: 10.1038/s41388-018-0625-1. Epub 2018 Dec 20.
Feller D, Kun J, Ruzsics I, Rapp J, Sarosi V, Kvell K, Helyes Z, Pongracz JE. Cigarette Smoke-Induced Pulmonary Inflammation Becomes Systemic by Circulating Extracellular Vesicles Containing Wnt5a and Inflammatory Cytokines. Front Immunol. 2018 Jul 25;9:1724. doi: 10.3389/fimmu.2018.01724. eCollection 2018.
Boda F, Banfai K, Garai K, Curticapean A, Berta L, Sipos E, Kvell K. Effect of Vipera ammodytes ammodytes Snake Venom on the Human Cytokine Network. Toxins (Basel). 2018 Jun 25;10(7). pii: E259. doi: 10.3390/toxins10070259.
Voros-Horvath Barbara; Zivkovic Pavo; Banfai Krisztina; Bovari-Biri Judit; Pongracz Judit; Balint Gabor; Pal; Szechenyi Aleksandar: Preparation and Characterization of ACE2 Receptor Inhibitor-Loaded Chitosan Hydrogels for Nasal Formulation to Reduce the Risk of COVID-19 Viral Infection. ACS Omega 2022
