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https://www.readbyqxmd.com/read/28064442/tumor-derived-spheroids-relevance-to-cancer-stem-cells-and-clinical-applications
#1
REVIEW
Tatsuya Ishiguro, Hirokazu Ohata, Ai Sato, Kaoru Yamawaki, Takayuki Enomoto, Koji Okamoto
Recently, many types of in vitro three-dimensional (3D) culture systems have been developed to recapitulate the in vivo growth conditions of cancer. The cancer 3D culture methods aim to preserve the biological characteristics of original tumors better than conventional two-dimensional (2D) monolayer cultures, and include tumor-derived organoids, tumor-derived spheroids, organotypic multicellular spheroids, and multicellular tumor spheroids. The 3D culture methods differ in terms of cancer cell sources, protocols for cell handling, and the required time intervals...
January 8, 2017: Cancer Science
https://www.readbyqxmd.com/read/28057213/a-new-molecular-mechanism-underlying-the-antitumor-effect-of-dna-methylation-inhibitors-via-an-antiviral-immune-response
#2
Y Saito, T Nakaoka, H Saito
Chromatin remodeling mediated by DNA methylation and histone modifications play critical roles in the transcriptional regulation of protein-coding genes, noncoding RNAs such as microRNAs, and endogenous retroviruses (ERVs). Many studies have shown that aberrant DNA methylation and histone modifications are associated with the initiation and progression of various malignancies. Epigenetic silencing of tumor suppressor genes in cancer is generally mediated by DNA hypermethylation of CpG island promoters and histone modifications such as histone deacetylation, methylation of histone H3 lysine 9 (H3K9), and trimethylation of H3K27...
2017: Advances in Protein Chemistry and Structural Biology
https://www.readbyqxmd.com/read/27993977/derivation-of-a-robust-mouse-mammary-organoid-system-for-studying-tissue-dynamics
#3
Paul R Jamieson, Johanna F Dekkers, Anne C Rios, Nai Yang Fu, Geoffrey J Lindeman, Jane E Visvader
Advances in stem cell research have enabled the generation of mini-organs or organoids that recapitulate phenotypic traits of the original biological specimen. Although organoids have been demonstrated for multiple organ systems, there are more limited options for studying mouse mammary gland formation in vitro Here we have built upon previously described culture assays to define culture conditions that enable the efficient generation of clonal organoid structures from single-sorted basal mammary epithelial cells (MECs)...
December 19, 2016: Development
https://www.readbyqxmd.com/read/27982115/generation-of-kidney-tubular-organoids-from-human-pluripotent-stem-cells
#4
Shintaro Yamaguchi, Ryuji Morizane, Koichiro Homma, Toshiaki Monkawa, Sayuri Suzuki, Shizuka Fujii, Muneaki Koda, Ken Hiratsuka, Maho Yamashita, Tadashi Yoshida, Shu Wakino, Koichi Hayashi, Junichi Sasaki, Shingo Hori, Hiroshi Itoh
Recent advances in stem cell research have resulted in methods to generate kidney organoids from human pluripotent stem cells (hPSCs), which contain cells of multiple lineages including nephron epithelial cells. Methods to purify specific types of cells from differentiated hPSCs, however, have not been established well. For bioengineering, cell transplantation, and disease modeling, it would be useful to establish those methods to obtain pure populations of specific types of kidney cells. Here, we report a simple two-step differentiation protocol to generate kidney tubular organoids from hPSCs with direct purification of KSP (kidney specific protein)-positive cells using anti-KSP antibody...
December 16, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27980341/induced-pluripotent-stem-cell-technology-a-decade-of-progress
#5
Yanhong Shi, Haruhisa Inoue, Joseph C Wu, Shinya Yamanaka
Since the advent of induced pluripotent stem cell (iPSC) technology a decade ago, enormous progress has been made in stem cell biology and regenerative medicine. Human iPSCs have been widely used for disease modelling, drug discovery and cell therapy development. Novel pathological mechanisms have been elucidated, new drugs originating from iPSC screens are in the pipeline and the first clinical trial using human iPSC-derived products has been initiated. In particular, the combination of human iPSC technology with recent developments in gene editing and 3D organoids makes iPSC-based platforms even more powerful in each area of their application, including precision medicine...
December 16, 2016: Nature Reviews. Drug Discovery
https://www.readbyqxmd.com/read/27973570/use-of-fluorescence-lifetime-imaging-microscopy-flim-as-a-timer-of-cell-cycle-s-phase
#6
Irina A Okkelman, Ruslan I Dmitriev, Tara Foley, Dmitri B Papkovsky
Incorporation of thymidine analogues in replicating DNA, coupled with antibody and fluorophore staining, allows analysis of cell proliferation, but is currently limited to monolayer cultures, fixed cells and end-point assays. We describe a simple microscopy imaging method for live real-time analysis of cell proliferation, S phase progression over several division cycles, effects of anti-proliferative drugs and other applications. It is based on the prominent (~ 1.7-fold) quenching of fluorescence lifetime of a common cell-permeable nuclear stain, Hoechst 33342 upon the incorporation of 5-bromo-2'-deoxyuridine (BrdU) in genomic DNA and detection by fluorescence lifetime imaging microscopy (FLIM)...
2016: PloS One
https://www.readbyqxmd.com/read/27960594/development-of-islet-organoids-from-h9-human-embryonic-stem-cells-in-biomimetic-3d-scaffolds
#7
Weiwei Wang, Sha Jin, Kaiming Ye
Success in the differentiating human embryonic stem cells (hESCs) into insulin-secreting β cells raises new hopes for diabetes treatment. In this work, we demonstrated the feasibility of developing islet organoids from hESCs within biomimetic 3D scaffolds. We showed that such a 3D microenvironment is critical to the generation of pancreatic endoderm and endocrine from hESCs. The organoids formed consisted of pancreatic α, β, δ, and pancreatic polypeptide (PP) cells. A high level co-expression of PDX1, NKX6...
December 13, 2016: Stem Cells and Development
https://www.readbyqxmd.com/read/27941077/post-translational-modification-of-%C3%AE-catenin-is-associated-with-pathogenic-fibroblastic-changes-in-bronchopulmonary-dysplasia
#8
Jennifer M S Sucre, Preethi Vijayaraj, Cody Aros, Dan Wilkinson, Manash Paul, Bruce Dunn, Susan H Guttentag, Brigitte N Gomperts
Bronchopulmonary dysplasia (BPD) is a common complication of premature birth. The histopathology of BPD is characterized by an arrest of alveolarization with fibroblast activation. The Wnt/β-catenin signaling pathway is important in early lung development. When Wnt signaling is active, phosphorylation of β-catenin by tyrosine kinases at activating sites, specifically at tyrosine 489 (Y489), correlates with nuclear localization of β-catenin. We examined fetal lung tissue, lung tissue from term newborns, and lung tissue from infants who died with BPD, and found nuclear β-catenin phosphorylatation at Y489 in epithelial and mesenchymal cells in fetal tissue and BPD tissue, but not the lungs of term infants...
December 9, 2016: American Journal of Physiology. Lung Cellular and Molecular Physiology
https://www.readbyqxmd.com/read/27909243/3d-culture-models-of-tissues-under-tension
#9
REVIEW
Jeroen Eyckmans, Christopher S Chen
Cells dynamically assemble and organize into complex tissues during development, and the resulting three-dimensional (3D) arrangement of cells and their surrounding extracellular matrix in turn feeds back to regulate cell and tissue function. Recent advances in engineered cultures of cells to model 3D tissues or organoids have begun to capture this dynamic reciprocity between form and function. Here, we describe the underlying principles that have advanced the field, focusing in particular on recent progress in using mechanical constraints to recapitulate the structure and function of musculoskeletal tissues...
January 1, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/27901089/a-silac-based-method-for-quantitative-proteomic-analysis-of-intestinal-organoids
#10
Alexis Gonneaud, Christine Jones, Naomie Turgeon, Dominique Lévesque, Claude Asselin, François Boudreau, François-Michel Boisvert
Organoids have the potential to bridge 3D cell culture to tissue physiology by providing a model resembling in vivo organs. Long-term growing organoids were first isolated from intestinal crypt cells and recreated the renewing intestinal epithelial niche. Since then, this technical breakthrough was applied to many other organs, including prostate, liver, kidney and pancreas. We describe here how to apply a SILAC-based quantitative proteomic approach to measure protein expression changes in intestinal organoids under different experimental conditions...
November 30, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27889058/mutations-in-reep6-cause-autosomal-recessive-retinitis-pigmentosa
#11
Gavin Arno, Smriti A Agrawal, Aiden Eblimit, James Bellingham, Mingchu Xu, Feng Wang, Christina Chakarova, David A Parfitt, Amelia Lane, Thomas Burgoyne, Sarah Hull, Keren J Carss, Alessia Fiorentino, Matthew J Hayes, Peter M Munro, Ralph Nicols, Nikolas Pontikos, Graham E Holder, Chinwe Asomugha, F Lucy Raymond, Anthony T Moore, Vincent Plagnol, Michel Michaelides, Alison J Hardcastle, Yumei Li, Catherine Cukras, Andrew R Webster, Michael E Cheetham, Rui Chen
Retinitis pigmentosa (RP) is the most frequent form of inherited retinal dystrophy. RP is genetically heterogeneous and the genes identified to date encode proteins involved in a wide range of functional pathways, including photoreceptor development, phototransduction, the retinoid cycle, cilia, and outer segment development. Here we report the identification of biallelic mutations in Receptor Expression Enhancer Protein 6 (REEP6) in seven individuals with autosomal-recessive RP from five unrelated families...
December 1, 2016: American Journal of Human Genetics
https://www.readbyqxmd.com/read/27882268/long-term-culture-of-intestinal-cell-progenitors-an-overview-of-their-development-application-and-associated-technologies
#12
REVIEW
Andrew J Hollins, Lee Parry
PURPOSE OF REVIEW: Long-term culture of adult progenitor cells in 3D is a recently emerging technology that inhabits the space between 2D cell lines and organ slice culture. RECENT FINDINGS: Adaptations to defined media components in the wake of advances in ES and iPS cell culture has led to the identification of conditions that maintained intestinal cell progenitors in culture. These conditions retain cellular heterogeneity of the normal or tumour tissue, and the cultures have been shown to be genetically stable, such that substantial biobanks are being created from patient derived material...
2016: Current Pathobiology Reports
https://www.readbyqxmd.com/read/27867006/intercellular-coupling-of-the-cell-cycle-and-circadian-clock-in-adult-stem-cell-culture
#13
Toru Matsu-Ura, Andrey Dovzhenok, Eitaro Aihara, Jill Rood, Hung Le, Yan Ren, Andrew E Rosselot, Tongli Zhang, Choogon Lee, Karl Obrietan, Marshall H Montrose, Sookkyung Lim, Sean R Moore, Christian I Hong
Circadian clock-gated cell division cycles are observed from cyanobacteria to mammals via intracellular molecular connections between these two oscillators. Here we demonstrate WNT-mediated intercellular coupling between the cell cycle and circadian clock in 3D murine intestinal organoids (enteroids). The circadian clock gates a population of cells with heterogeneous cell-cycle times that emerge as 12-hr synchronized cell division cycles. Remarkably, we observe reduced-amplitude oscillations of circadian rhythms in intestinal stem cells and progenitor cells, indicating an intercellular signal arising from differentiated cells governing circadian clock-dependent synchronized cell division cycles...
December 1, 2016: Molecular Cell
https://www.readbyqxmd.com/read/27859262/functional-and-prognostic-significance-of-the-genomic-amplification-of-frizzled-6-fzd6-in-breast-cancer
#14
Gabriele Corda, Gianluca Sala, Rossano Lattanzio, Manuela Iezzi, Michele Sallese, Giorgia Fragassi, Alessia Lamolinara, Hasan Mirza, Daniela Barcaroli, Sibylle Ermler, Elisabete Silva, Hemad Yasaei, Robert F Newbold, Paola Vagnarelli, Marcella Mottolese, Pier Giorgio Natali, Letizia Perracchio, Jelmar Quist, Anita Grigoriadis, Pierfrancesco Marra, Andrew N Tutt, Mauro Piantelli, Stefano Iacobelli, Vincenzo De Laurenzi, Arturo Sala
Frizzled receptors mediate Wnt ligand signalling, which is crucially involved in regulating tissue development and differentiation, and is often deregulated in cancer. In this study, we found that the gene encoding the Wnt receptor frizzled 6 (FZD6) is frequently amplified in breast cancer, with an increased incidence in the triple-negative breast cancer (TNBC) subtype. Ablation of FZD6 expression in mammary cancer cell lines: (1) inhibited motility and invasion; (2) induced a more symmetrical shape of organoid three-dimensional cultures; and (3) inhibited bone and liver metastasis in vivo...
November 9, 2016: Journal of Pathology
https://www.readbyqxmd.com/read/27844287/modeling-of-autism-using-organoid-technology
#15
REVIEW
Hwan Choi, Juhyun Song, Guiyeon Park, Jongpil Kim
Autism is a neurodevelopmental disease caused by multiple mutations during development. However, a suitable disease model to study the molecular pathway of disease onset and progression is not available. Although many studies have used human stem cells such as induced pluripotent stem cells and embryonic stem cells to investigate the disease pathogenesis, these stem cell techniques are limited in their abilities to study the pathology and mechanism of pathogenesis of neurodevelopmental diseases such as autism...
November 14, 2016: Molecular Neurobiology
https://www.readbyqxmd.com/read/27827428/pge2-is-a-direct-and-robust-mediator-of-anion-fluid-secretion-by-human-intestinal-epithelial-cells
#16
Satoru Fujii, Kohei Suzuki, Ami Kawamoto, Fumiaki Ishibashi, Toru Nakata, Tatsuro Murano, Go Ito, Hiromichi Shimizu, Tomohiro Mizutani, Shigeru Oshima, Kiichiro Tsuchiya, Tetsuya Nakamura, Akihiro Araki, Kazuo Ohtsuka, Ryuichi Okamoto, Mamoru Watanabe
Intestinal epithelial cells (IECs) play an indispensable role in maintaining body fluid balance partly through their ability to regulate anion/fluid secretion. Yet in various inflammatory gastrointestinal diseases, over-secretion of anions results in symptoms such as severe diarrhoea. Endogenous mediators, such as vasoactive intestinal peptide or prostaglandin E2 (PGE2), regulate intestinal anion/fluid secretion, but their direct effect on purified human IECs has never been described in detail. Based on a previously described intestinal organoid swelling model, we established a 3D-scanner-assisted quantification method to evaluate the anion/fluid secretory response of cultured human IECs...
November 9, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27815996/human-airway-organoid-engineering-as-a-step-toward-lung-regeneration-and-disease-modeling
#17
Qi Tan, Kyoung Moo Choi, Delphine Sicard, Daniel J Tschumperlin
Organoids represent both a potentially powerful tool for the study cell-cell interactions within tissue-like environments, and a platform for tissue regenerative approaches. The development of lung tissue-like organoids from human adult-derived cells has not previously been reported. Here we combined human adult primary bronchial epithelial cells, lung fibroblasts, and lung microvascular endothelial cells in supportive 3D culture conditions to generate airway organoids. We demonstrate that randomly-seeded mixed cell populations undergo rapid condensation and self-organization into discrete epithelial and endothelial structures that are mechanically robust and stable during long term culture...
January 2017: Biomaterials
https://www.readbyqxmd.com/read/27796955/a-3d-fibroblast-epithelium-co-culture-model-for-understanding-microenvironmental-role-in-branching-morphogenesis-of-the-mammary-gland
#18
Zuzana Koledova, Pengfei Lu
The mammary gland consists of numerous tissue compartments, including mammary epithelium, an array of stromal cells, and the extracellular matrix (ECM). Bidirectional interactions between the epithelium and its surrounding stroma are essential for proper mammary gland development and homeostasis, whereas their deregulation leads to developmental abnormalities and cancer. To study the relationships between the epithelium and the stroma, development of models that could recapitulate essential aspects of these interacting systems in vitro has become necessary...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27787775/the-three-dimensional-culture-of-epithelial-organoids-derived-from-embryonic-chicken-intestine
#19
Malgorzata Pierzchalska, Malgorzata Panek, Malgorzata Czyrnek, Maja Grabacka
The intestinal epithelium isolated from chicken embryos in last 3 days of development can be used to establish the 3D culture of intestinal organoids. When fragments of epithelial tissue released by incubation with EGTA (2.5 mM, 2 h) are embedded in Matrigel matrix on cell culture inserts the formation of empty spheres covered by epithelial cells is observed in first 24 h of culture. The growth and survival of organoids are supported by the addition of R-spondin 1, Noggin, and prostaglandin E2 to the culture medium...
October 28, 2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27783065/the-promises-and-challenges-of-human-brain-organoids-as-models-of-neuropsychiatric-disease
#20
Giorgia Quadrato, Juliana Brown, Paola Arlotta
Neuropsychiatric disorders such as autism spectrum disorder (ASD), schizophrenia (SCZ) and bipolar disorder (BPD) are of great societal and medical importance, but the complexity of these diseases and the challenges of modeling the development and function of the human brain have made these disorders difficult to study experimentally. The recent development of 3D brain organoids derived from human pluripotent stem cells offers a promising approach for investigating the phenotypic underpinnings of these highly polygenic disorders and for understanding the contribution of individual risk variants and complex genetic background to human pathology...
October 26, 2016: Nature Medicine
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