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3D organoid

Xiao Yu, Ashley M Fuller, Richard Blackmon, Melissa A Troester, Amy L Oldenburg
The ability to assess toxicant exposures of three-dimensional (3D) in vitro mammary models that recapitulate the tissue microenvironment can aid in our understanding of environmental exposure risk over time. Longitudinal studies of 3D model systems, however, are cumbersome and suffer from a lack of high-throughput toxicological assays. In this study, we establish a noninvasive and label-free optical coherence tomography (OCT)-based imaging platform for tracking exposure-response relationships in 3D human mammary epithelial organoid models...
November 10, 2017: Toxicological Sciences: An Official Journal of the Society of Toxicology
Richard J Maunder, Matthew G Baron, Stewart F Owen, Awadhesh N Jha
The primary culture of fish gill cells can provide functional, cell diverse, model in vitro platforms able to tolerate an aqueous exposure analogous to in vivo tissues. The utility of such models could be extended to a variety of longer term exposure scenarios if a method could be established to extend culture viability when exposed to water for longer periods. Here we report findings of a series of experiments to establish increased longevity, as monitored by culture transepithelial electrical resistance (TEER) and concurrent histological developments...
November 11, 2017: Ecotoxicology
Eva Tomaskovic-Crook, Jeremy M Crook
Human brain organoids provide opportunities to produce three-dimensional (3D) brain-like tissues for biomedical research and translational drug discovery, toxicology, and tissue replacement. Here we describe a protocol for rapid and defined induction of brain organoids from human induced pluripotent stem cells (iPSCs), using commercially available culture and differentiation media and a cheap, easy to handle and clinically approved semisynthetic hydrogel. Importantly, the methodology is uncomplicated, well-defined, and reliable for reproducible and scalable organoid generation, and amendable to principles of current good laboratory practice (cGLP), with the potential for prospective adaptation to current good manufacturing practice (cGMP) toward clinical compliance...
November 9, 2017: Methods in Molecular Biology
Fakhar Singhera, Emily Cooper, Louis Scampavia, Timothy Spicer
Biomedical translational research has relied on two dimensional (2D) cell cultures for drug discovery over the decades, requiring cells to grow on a flat surface which does not always accurately model in vivo biological states. Three dimensional (3D) cell cultures, also known as 3D spheroids or organoids, grow as cellular tissues that are more physiologically relevant especially with respect to emulating cancer tumor-like structures [1]. While attractive, current methods for generating 3D spheroids has yet to replace 2D culturing methods used for drug discovery efforts that employ high-throughput screening (HTS), having limitations with scalability, reproducibility, and compatibility predominantly associated with conventional microtiter plate usage...
January 15, 2018: Talanta
Tânia Rodrigues, Banani Kundu, Joana Silva-Correia, S C Kundu, Joaquim M Oliveira, Rui L Reis, Vitor M Correlo
Cancer is a leading cause of mortality and morbidity worldwide. Around 90% of deaths are caused by metastasis and just 10% by primary tumor. The advancement of treatment approaches is not at the same rhythm of the disease; making cancer a focal target of biomedical research. To enhance the understanding and promts the therapeutic delivery; concepts of tissue engineering are applied in the development of in vitro models that can bridge between 2D cell culture and animal models, mimicking tissue microenvironment...
October 30, 2017: Pharmacology & Therapeutics
Haristi Gaitantzi, Priska Hakenberg, Jannick Theobald, Hagen Heinlein, Chen Cai, Steffan Loff, Stefan Wölfl, Matthias P Ebert, Katja Breitkopf-Heinlein, Ulrike Subotic
OBJECTIVES: Di(2-ethylhexyl) phthalate (DEHP) is a plasticizer used in many polyvinylchloride medical devices and is washed out easily. Thereby critically ill infants can become exposed to DEHP concentrations significantly exceeding the recommended threshold. We suspect DEHP to play an important role in the development of intestinal failure associated liver disease. The aim of this study was therefore, to determine the direct influence of DEHP on different liver cell types. METHODS: HepG2, human upcyte hepatocytes, primary murine hepatocytes, LX-2, human upcyte hepatic stellate cells and liver organoids were cultured with DEHP (0...
October 31, 2017: Journal of Pediatric Gastroenterology and Nutrition
Yusuke Yanagi, Koichi Nakayama, Tomoaki Taguchi, Shin Enosawa, Tadashi Tamura, Koichiro Yoshimaru, Toshiharu Matsuura, Makoto Hayashida, Kenichi Kohashi, Yoshinao Oda, Takayoshi Yamaza, Eiji Kobayashi
Cell-based therapy has been proposed as an alternative to orthotopic liver transplantation. The novel transplantation of an in vitro-generated liver bud might have therapeutic potential. In vivo and ex vivo methods for growing a liver bud are essential for paving the way for the clinical translation of liver bud transplantation. We herein report a novel transplantation method for liver buds that are grown in vivo involving orthotopic transplantation on the transected parenchyma of the liver, which showed long engraftment and marked growth in comparison to heterotopic transplantation...
October 26, 2017: Scientific Reports
D P Ivanov, A M Grabowska
Three-dimensional (3D) in vitro microphysiological cultures, such as spheroids and organoids, promise increased patient relevance and therapeutic predictivity compared with reductionist cell monolayers. However, high-throughput characterization techniques for 3D models are currently limited to simplistic live/dead assays. By sectioning and staining in vitro microtissues, researchers can examine their structure; detect DNA, RNA, and protein targets; and visualize them at the level of single cells. The morphological examination and immunochemistry staining for in vitro cultures has historically been done in a laborious manner involving testing one set of cultures at a time...
October 1, 2017: SLAS Discovery
Angela W Xie, Bernard Y K Binder, Andrew S Khalil, Samantha K Schmitt, Hunter J Johnson, Nicholas A Zacharias, William L Murphy
Stem cell-derived organoids and other 3D microtissues offer enormous potential as models for drug screening, disease modeling, and regenerative medicine. Formation of stem/progenitor cell aggregates is common in biomanufacturing processes and critical to many organoid approaches. However, reproducibility of current protocols is limited by reliance on poorly controlled processes (e.g., spontaneous aggregation). Little is known about the effects of aggregation parameters on cell behavior, which may have implications for the production of cell aggregates and organoids...
October 25, 2017: Scientific Reports
Akiko Mito, Yukiko Nakano, Takako Saitoh, Sabine S S Gouraud, Yoshiki Yamaguchi, Toshiro Sato, Nobuo Sasaki, Kyoko Kojima-Aikawa
Zymogen granule protein 16 (ZG16p) is a soluble lectin that binds to both mannose and heparin/heparan sulfate. It is highly expressed in the human digestive tract and is secreted into the mucus. In this study, we investigated the effect of ZG16p on the proliferation of human colorectal cancer cells. Overexpression of ZG16p in Caco-2 cells decreased cell growth. Recombinant ZG16p markedly inhibited proliferation of Caco-2, LS174T, HCT116 and HCT15 cells. Caco-2 cell growth was not inhibited by two mutated ZG16p proteins, D151A and M5 (K36A, R37A, R53A, R55A and R79A) lacking mannose- and heparin-binding activities, respectively...
October 23, 2017: Glycobiology
Jesús Gómez-Escudero, Vanessa Moreno, Mara Martín-Alonso, M Victoria Hernández de Riquer, Tamar Feinberg, Ángel Colmenar, Enrique Calvo, Emilio Camafeita, Fernando Martínez, Menno J Oudhoff, Stephen J Weiss, Alicia G Arroyo
Cadherin-based intercellular adhesions are essential players in epithelial homeostasis, but their dynamic regulation during tissue morphogenesis and remodeling remain largely undefined. Herein, we characterize an unexpected role for the membrane-anchored metalloproteinase MT2-MMP in regulating epithelial cell quiescence. Following co-immunoprecipitation and mass spectrometry, the MT2-MMP cytosolic tail was found to interact with the zonula occludens protein-1 (ZO-1) at the apical junctions of polarized epithelial cells...
October 23, 2017: Journal of Cell Science
Amy Q Lu, Colin J Barnstable
Embryonic stem cell (ESC) differentiation can be used to model development and to produce transplantable cells of the desired phenotype. ESCs can reproducibly generate retinal cells but the derivation of photoreceptor precursors is variable and depends on an array of exogenous factors and intrinsic cell-cell interactions. In this work, we have defined the use of exogenous signaling factors, dissociation, and adherent versus 3-dimensional (3D) conditions on the derivation of retinal cells from pluripotent mouse ESCs...
October 18, 2017: Stem Cell Reviews
Wendy Béguelin, Martín A Rivas, María T Calvo Fernández, Matt Teater, Alberto Purwada, David Redmond, Hao Shen, Matt F Challman, Olivier Elemento, Ankur Singh, Ari M Melnick
The EZH2 histone methyltransferase is required for B cells to form germinal centers (GC). Here we show that EZH2 mediates GC formation through repression of cyclin-dependent kinase inhibitor CDKN1A (p21(Cip1)). Deletion of Cdkn1a rescues the GC reaction in Ezh2 (-/-) mice. Using a 3D B cell follicular organoid system that mimics the GC reaction, we show that depletion of EZH2 suppresses G1 to S phase transition of GC B cells in a Cdkn1a-dependent manner. GC B cells of Cdkn1a (-/-) Ezh2 (-/-) mice have high levels of phospho-Rb, indicating that loss of Cdkn1a enables progression of cell cycle...
October 12, 2017: Nature Communications
Tatsuya Usui, Masashi Sakurai, Shimpei Nishikawa, Koji Umata, Yuki Nemoto, Tomoya Haraguchi, Kazuhito Itamoto, Takuya Mizuno, Shunsuke Noguchi, Takashi Mori, Satomi Iwai, Takayuki Nakagawa, Hideyuki Yamawaki, Takashi Ohama, Koichi Sato
Dog spontaneously develops prostate cancer (PC) like human. Since most dogs with PC have poor prognosis, they are expected to become a translational model for human advanced PC. Stem cell-derived 3D organoid culture could recapitulate organ structures and physiology. Using patient tissues, human PC organoid culture system was established. Recent study showed that urine cells also possess the characteristic of stem cells. However, the urine cell-derived PC organoids have never been produced. We therefore generated PC organoids using the dog urine samples...
October 10, 2017: Cancer Science
Dwaipayan Adhya, Emily Annuario, Madeline A Lancaster, Jack Price, Simon Baron-Cohen, Deepak P Srivastava
Steroids have an important role in growth, development, sexual differentiation and reproduction. All four classes of steroids, androgens, estrogens, progestogens and glucocorticoids, have varying effects on the brain. Androgens and estrogens are involved in the sexual differentiation of the brain, and also influence cognition. Progestogens such as progesterone and its metabolites have been shown to be involved in neuroprotection, although their protective effects are timing dependent. Glucocorticoids are linked with stress and memory performance, also in a dose- and time-dependent manner...
October 12, 2017: Journal of Neuroendocrinology
Elena Garreta, Federico González, Núria Montserrat
Kidney morphogenesis and patterning have been extensively studied in animal models such as the mouse and zebrafish. These seminal studies have been key to define the molecular mechanisms underlying this complex multistep process. Based on this knowledge, the last 3 years have witnessed the development of a cohort of protocols allowing efficient differentiation of human pluripotent stem cells (hPSCs) towards defined kidney progenitor populations using two-dimensional (2D) culture systems or through generating organoids...
October 7, 2017: Nephron
Aslam Abbasi Akhtar, Samuel Sances, Robert Barrett, Joshua J Breunig
PURPOSE OF REVIEW: The modeling of biological processes in vitro provides an important tool to better understand mechanisms of development and disease, allowing for the rapid testing of therapeutics. However, a critical constraint in traditional monolayer culture systems is the absence of the multicellularity, spatial organization, and overall microenvironment present in vivo. This limitation has resulted in numerous therapeutics showing efficacy in vitro, but failing in patient trials...
June 2017: Current Stem Cell Reports
Nikolce Gjorevski, Matthias P Lutolf
Growing cells within an extracellular matrix-like 3D gel is required for, or can improve, the growth of many cell types ex vivo. Here, we describe a protocol for the generation of well-defined matrices for the culture of intestinal stem cells (ISCs) and intestinal organoids. These matrices comprise a poly(ethylene glycol) (PEG) hydrogel backbone functionalized with minimal adhesion cues including RGD (Arg-Gly-Asp), which is sufficient for ISC expansion, and laminin-111, which is required for organoid formation...
November 2017: Nature Protocols
Anastasia Tsakmaki, Patricia Fonseca Pedro, Gavin A Bewick
The advent of near physiological organoid technology has produced a step change in our understanding of stem cells and has provided the research community with a powerful new cell based tool to model human physiology and disease. We review the pros and cons of intestinal organoid culture systems. The molecular and genetic tools to manipulate them and how they are being used to answer fundamental questions in metabolic research, including the function of enteroendocrine cells in health and disease.
September 29, 2017: Current Opinion in Pharmacology
Victor Tostivint, Claire Racaud-Sultan, Mathieu Roumiguié, Michel Soulié, Xavier Gamé, Jean-Baptiste Beauval
Despite new therapeutics options, Prostate Cancer (PCa) remains a public health challenge because of its high incidence and mortality. Limits in PCa research come from the lack of in vitro and in vivo models that mimic the human disease. Currently, 2D in vitro tissue culture models of PCa are widely used but they present numerous limits. They do not reproduce cellular morphology, tissue architecture, inter-patients and intratumor heterogeneity. Furthermore, they lack two key components of PCa tumors, the tumoral microenvironment and the cancer stem cells...
September 26, 2017: La Presse Médicale
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