The Experts below are selected from a list of 74208 Experts worldwide ranked by ideXlab platform
Albert De La Chapelle - One of the best experts on this subject based on the ideXlab platform.
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Primary Cell Culture systems for human thyroid studies
Thyroid, 2016Co-Authors: Yanqiang Wang, John E Phay, Rulong Shen, Natalia S Pellegata, Motoyasu Saji, Matthew D Ringel, Albert De La ChapelleAbstract:Background: Cell models are key instruments for in vitro studies of the thyroid. Permanent thyroid Cell lines that are widely used in laboratory research typically originate from tumors. For many purposes, it is desirable to compare tumor Cells with Cells originating from normal tissue. However, such Cultures grow slowly, have a highly limited life-span, and are known to lose their thyroid characteristics. The aim of the present study was to type coding and noncoding thyroid markers in different Culture systems in an attempt to determine the optimal conditions for in vitro experimentation. Methods: Human Primary thyroid Cells were isolated from histologically non-tumorous tissues. Two alternative media (6H and h7H) were used. The morphology and behavior of the ensuing monolayer (two-dimensional) Cultures was monitored by microscopy. The expression of key thyroid-related genes (n = 9) was monitored by reverse transcription polymerase chain reaction on days 8, 21, and 43 after initiation. As a pilot study, ...
Thomas F Meyer - One of the best experts on this subject based on the ideXlab platform.
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a novel human gastric Primary Cell Culture system for modelling helicobacter pylori infection in vitro
Gut, 2016Co-Authors: Philipp Schlaermann, Benjamin Toelle, Hilmar Berger, Sven Schmidt, Matthias Glanemann, Jurgen Ordemann, Sina Bartfeld, Hans-joachim Mollenkopf, Thomas F MeyerAbstract:Background and aims Helicobacter pylori is the causative agent of gastric diseases and the main risk factor in the development of gastric adenocarcinoma. In vitro studies with this bacterial pathogen largely rely on the use of transformed Cell lines as infection model. However, this approach is intrinsically artificial and especially inappropriate when it comes to investigating the mechanisms of cancerogenesis. Moreover, common Cell lines are often defective in crucial signalling pathways relevant to infection and cancer. A long-lived Primary Cell system would be preferable in order to better approximate the human in vivo situation. Methods Gastric glands were isolated from healthy human stomach tissue and grown in Matrigel containing media supplemented with various growth factors, developmental regulators and apoptosis inhibitors to generate long-lasting normal epithelial Cell Cultures. Results Culture conditions were developed which support the formation and quasi-indefinite growth of three dimensional (3D) spheroids derived from various sites of the human stomach. Spheroids could be differentiated to gastric organoids after withdrawal of Wnt3A and R-spondin1 from the medium. The 3D Cultures exhibit typical morphological features of human stomach tissue. Transfer of sheared spheroids into 2D Culture led to the formation of dense planar Cultures of polarised epithelial Cells serving as a suitable in vitro model of H. pylori infection. Conclusions A robust and quasi-immortal 3D organoid model has been established, which is considered instrumental for future research aimed to understand the underlying mechanisms of infection, mucosal immunity and cancer of the human stomach.
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abstract 2028 a novel human gastric Primary Cell Culture system for modelling helicobacter pylori infection in vitro
Cancer Research, 2014Co-Authors: Philipp Schlaermann, Thomas F MeyerAbstract:Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA The significance of bacterial infections in the initiation of human cancers is increasingly being recognized. A well established example is Helicobacter pylori, a causative agent of gastric adenocarcinoma. Current research largely relies on the use of transformed human Cell lines as in vitro models of these infections. Obviously, this concept is largely artificial and especially inappropriate when it comes to investigations on the mechanisms of cancerogenesis. A Primary Cell system would be preferable in order to approximate the in vivo situation, however, previous attempts to establish such models are hampered by the limited growth and reproducibility of normal human Primary Cell Cultures. Here, we report on the establishment of specific long-term Culture conditions that allow the formation of 3-dimensional human gastric organoids with epithelial domains that have the capacity to self-renew and can be maintained in Culture indefinitely. Interestingly, these 3-dimensional structures can be transferred into 2-dimensional Primary Cell layers, which provide ideal conditions to study infections in vitro. We expect that our novel human gastric Primary Cell model will facilitate investigations on the mechanisms underlying infection and is suited to open up avenues of approach to investigations on the initial processes of Cellular transformation from normal to a transformed Cell stages including the tumor-promoting capacity of the gastric pathogen H. pylori. Citation Format: Philipp Schlaermann, Thomas F. Meyer. A novel human gastric Primary Cell Culture system for modelling Helicobacter pylori infection in vitro. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2028. doi:10.1158/1538-7445.AM2014-2028
Yanqiang Wang - One of the best experts on this subject based on the ideXlab platform.
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Primary Cell Culture systems for human thyroid studies
Thyroid, 2016Co-Authors: Yanqiang Wang, John E Phay, Rulong Shen, Natalia S Pellegata, Motoyasu Saji, Matthew D Ringel, Albert De La ChapelleAbstract:Background: Cell models are key instruments for in vitro studies of the thyroid. Permanent thyroid Cell lines that are widely used in laboratory research typically originate from tumors. For many purposes, it is desirable to compare tumor Cells with Cells originating from normal tissue. However, such Cultures grow slowly, have a highly limited life-span, and are known to lose their thyroid characteristics. The aim of the present study was to type coding and noncoding thyroid markers in different Culture systems in an attempt to determine the optimal conditions for in vitro experimentation. Methods: Human Primary thyroid Cells were isolated from histologically non-tumorous tissues. Two alternative media (6H and h7H) were used. The morphology and behavior of the ensuing monolayer (two-dimensional) Cultures was monitored by microscopy. The expression of key thyroid-related genes (n = 9) was monitored by reverse transcription polymerase chain reaction on days 8, 21, and 43 after initiation. As a pilot study, ...
Shuo Lin - One of the best experts on this subject based on the ideXlab platform.
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high throughput screening for bioactive molecules using Primary Cell Culture of transgenic zebrafish embryos
Cell Reports, 2012Co-Authors: Haigen Huang, Anne Lindgren, Ningai Liu, Shuo LinAbstract:Transgenic zebrafish embryos expressing tissue-specific green fluorescent protein (GFP) can provide an unlimited supply of Primary embryonic Cells. Agents that promote the differentiation of these Cells may be beneficial for therapeutics. We report a high-throughput approach for screening small molecules that regulate Cell differentiation using lineage-specific GFP transgenic zebrafish embryonic Cells. After validating several known regulators of the differentiation of endothelial and other Cell types, we performed a screen for proangiogenic molecules using undifferentiated Primary Cells from flk1-GFP transgenic zebrafish embryos. Cells were grown in 384-well plates with 12,128 individual small molecules, and GFP expression was analyzed by means of an automated imaging system, which allowed us to screen thousands of compounds weekly. As a result, 23 molecules were confirmed to enhance angiogenesis, and 11 of them were validated to promote the proliferation of mammalian human umbilical vascular endothelial Cells and induce Flk1+ Cells from murine embryonic stem Cells. We demonstrated the general applicability of this strategy by analyzing additional Cell lineages using zebrafish expressing GFP in pancreatic, cardiac, and dopaminergic Cells.
Christoph A Reinhardt - One of the best experts on this subject based on the ideXlab platform.
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neurodevelopmental toxicity in vitro Primary Cell Culture models for screening and risk assessment
Reproductive Toxicology, 1993Co-Authors: Christoph A ReinhardtAbstract:Abstract Robust models for the evaluation of developmental toxicity are briefly reviewed with emphasis on embryonic brain and retina Cells in vitro. Organ slice and aggregate Cultures under constant gyratory movement as well as high Cell density monolayer (“micromass”) Cultures are considered as robust models. An in vitro model using high Cell density monolayer and re-aggregated Cells isolated from embryonic chick brain (ED 6) is presented. Cell development and differentiation of the astrocytes and nerve Cells are monitored by marker proteins and cytotoxicity was quantified by neutral red uptake and protein content. Four human teratogens, six possible human teratogens and six unlikely human teratogens were tested in brain and retina Cells for their cytotoxic and morphologic effect. All 16 substances were classified correctly except the neurotoxicants MPTP and MPP+, both of which are strong dopaminergic toxicants in vitro as well as in humans and are therefore proposed to be classified as human neuroteratogens. Preliminary data on the lowest effect levels of four potential neurotoxicants (cadmium chloride, Ara-C, Phenytoin, MPTP) in chick brain aggregate Cultures correlate suprisingly well with known toxic human plasma levels. Further validation has to be undertaken to confirm these promising results. A battery of such robust in vitro models is proposed that could cover neurodevelopmental toxicity of drugs and chemicals for screening and risk assessment purposes.