scholarly journals A Novel High-Throughput Screening Platform Identifies Itaconate Derivatives from Marine Penicillium antarcticum as Inhibitors of Mesenchymal Stem Cell Differentiation

Marine Drugs ◽  
2020 ◽  
Vol 18 (4) ◽  
pp. 192
Author(s):  
Pietro Marchese ◽  
Nipun Mahajan ◽  
Enda O’Connell ◽  
Howard Fearnhead ◽  
Maria Tuohy ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Differentiation ◽  
Small Molecules ◽  
High Throughput ◽  
High Throughput Screening ◽  
Chondrogenic Differentiation ◽  
Stem Cell Differentiation ◽  
Ectopic Calcification ◽  
Tissue Calcification ◽  
Screening Platform

Worldwide diffused diseases such as osteoarthritis, atherosclerosis or chronic kidney disease are associated with a tissue calcification process which may involve unexpected local stem cell differentiation. Current pharmacological treatments for such musculoskeletal conditions are weakly effective, sometimes extremely expensive and often absent. The potential to develop new therapies is represented by the discovery of small molecules modulating resident progenitor cell differentiation to prevent aberrant tissue calcification. The marine environment is a rich reserve of compounds with pharmaceutical potential and many novel molecules are isolated from macro and microorganisms annually. The potential of small molecules synthetized by marine filamentous fungi to influence the osteogenic and chondrogenic differentiation of human mesenchymal stem/stromal cells (hMSCs) was investigated using a novel, high-throughput automated screening platform. Metabolites synthetized by the marine-derived fungus Penicillium antarcticum were evaluated on the platform. Itaconic acid derivatives were identified as inhibitors of calcium elaboration into the matrix of osteogenically differentiated hMSCs and also inhibited hMSC chondrogenic differentiation, highlighting their capacity to impair ectopic calcification. Bioactive small molecule discovery is critical to address ectopic tissue calcification and the use of biologically relevant assays to identify naturally occurring metabolites from marine sources represents a strategy that can contribute to this effort.

The use of chondrogenic differentiation drugs to induce stem cell differentiation using double bead microsphere structure

Biomaterials ◽  
2008 ◽  
Vol 29 (16) ◽  
pp. 2490-2500 ◽  
Author(s):  
Kyeongsoon Park ◽  
Ji Sun Park ◽  
Dae Gyun Woo ◽  
Han Na Yang ◽  
Hyung-Min Chung ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Differentiation ◽  
Chondrogenic Differentiation ◽  
Stem Cell Differentiation

scholarly journals A high-throughput screening platform for pigment regulating agents using pluripotent stem cell derived melanocytes

2020 ◽  
Author(s):  
Valentin Parat ◽  
Brigitte Onteniente ◽  
Julien Maruotti
Keyword(s):  
Stem Cell ◽  
High Throughput ◽  
High Throughput Screening ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Light Scatter ◽  
Chemical Treatments ◽  
Human Melanocytes ◽  
Screening Platform ◽  
Induced Pluripotent

AbstractIn this study, we describe a simple and straight-forward assay using induced pluripotent stem cell derived melanocytes and high-throughput flow cytometry, to screen and identify pigment regulating agents. The assays is based on the correlation between forward light-scatter characteristics and melanin content, with pigmented cells displaying high light absorption/low forward light-scatter, while the opposite is true for lowly pigmented melanocytes, as a result of genetic background or chemical treatments. Orthogonal validation is then performed by regular melanin quantification. Such approach was validated using a set of 80 small molecules, and yielded a confirmed hit. The assay described in this study may prove a useful tool to identify modulators of melanogenesis in human melanocytes.

scholarly journals Monitoring stem cell differentiation using Raman microspectroscopy: chondrogenic differentiation, towards cartilage formation

The Analyst ◽  
2021 ◽  
Vol 146 (1) ◽  
pp. 322-337
Author(s):  
Francesca Ravera ◽  
Esen Efeoglu ◽  
Hugh J. Byrne
Keyword(s):  
Stem Cells ◽  
Extracellular Matrix ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Cell Differentiation ◽  
Chondrogenic Differentiation ◽  
Stem Cell Differentiation ◽  
Raman Microspectroscopy ◽  
Cartilage Formation ◽  
Matrix Evolution

Raman microspectroscopy is employed to monitor the differentiation of mesenchymal stem cells to chondrocytes, from subcellular to extracellular matrix evolution.

scholarly journals High-Throughput Screening Platform Identifies Small Molecules That Prevent Sequestration ofPlasmodium falciparum–Infected Erythrocytes

2014 ◽  
Vol 211 (7) ◽  
pp. 1134-1143 ◽  
Author(s):  
Justin Gullingsrud ◽  
Neta Milman ◽  
Tracy Saveria ◽  
Olga Chesnokov ◽  
Kathryn Williamson ◽  
...  
Keyword(s):  
Small Molecules ◽  
High Throughput ◽  
High Throughput Screening ◽  
Ofplasmodium Falciparum ◽  
Screening Platform

scholarly journals LAA and CoCl2 Pretreated Exosomes Enhance Chondrogenic Differentiation of Mesenchymal Stem Cell

2021 ◽  
Author(s):  
Anggraini Barlian ◽  
Rizka Musdalifah Amsar ◽  
Salindri Prawitasari ◽  
Christofora Hanny Wijaya ◽  
Ika Dewi Ana ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Cell Differentiation ◽  
Mesenchymal Stem Cell ◽  
Chondrogenic Differentiation ◽  
Stem Cell Differentiation ◽  
Human Umbilical Cord ◽  
Hypoxic Conditions ◽  
Transmission Electron

Abstract BackgroundCells produce extracellular vesicles, such as exosomes and microvesicles, which are used for intracellular communication. Cell-free therapies could be enhanced by using mesenchymal stem cell-derived exosomes. Preconditioning parental cells affects the properties of their exosomes. This study aimed to investigate the role of L-ascorbic acid (LAA) and CoCl2 in the exosomes produced by human Wharton’s jelly mesenchymal stem cells (hWJ MSC) and its potential to induce chondrogenic differentiation of stem cells was also studied.. MethodThe cells were obtained from umbilical cords and characterized based on mesenchymal stem cell criteria. The cells were cultured in a serum-free medium containing LAA and CoCl2. Exosomes produced by the cells were isolated and their morphology observed with Transmission Electron Microscopy. The presence of CD 63 was confirmed using ELISA. The particle size distribution and exosome concentration were analyzed with Nanoparticle Tracking Analysis (NTA). The ability of exosomes to induce stem cell differentiation into chondrocytes was investigated using the Alcian blue assay and immunocytochemistry.ResultsStem cells were successfully isolated from the human umbilical cord. The cells can differentiate into adipocytes, chondrocytes, and osteocytes. Flowcytometry analysis showed the specific surface marker of mesenchymal stem cells. Exosomes isolated from pretreatment cells showed round-shaped morphology and confirmed the presence of CD 63. NTA analysis revealed that pretreatment of cells with LAA increases exosome yields. LAA supplementation in cell medium under hypoxic conditions induced by CoCl2 produces exosomes that can induce the chondrogeic differentiation of stem cells, confirmed by the presence of glycosaminoglycan and collagen type 2.ConclusionExosomes produced by preconditioning hWJ-MSC with LAA in hypoxic conditions have the potential to enhance human Wharton Jelly stem cell differentiation into chondrocytes.

scholarly journals High-throughput organoid screening enables engineering of intestinal epithelial composition

2020 ◽  
Author(s):  
Benjamin E. Mead ◽  
Kazuki Hattori ◽  
Lauren Levy ◽  
Marko Vukovic ◽  
Daphne Sze ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cell Differentiation ◽  
High Throughput ◽  
Intestinal Epithelium ◽  
Paneth Cell ◽  
Stem Cell Differentiation ◽  
Biological Targets ◽  
Small Intestinal

SummaryBarrier tissue epithelia play an essential role in maintaining organismal homeostasis, and changes in their cellular composition have been observed in multiple human diseases. Within the small intestinal epithelium, adult stem cells integrate diverse signals to regulate regeneration and differentiation, thereby establishing overall cellularity. Accordingly, directing stem cell differentiation could provide a tractable approach to alter the abundance or quality of specialized cells of the small intestinal epithelium, including the secretory Paneth, goblet, and enteroendocrine populations. Yet, to date, there has been a lack of suitable tools and rigorous approaches to identify biological targets and pharmacological agents that can modify epithelial composition to enable causal testing of disease-associated changes with novel therapeutic candidates. To empower the search for epithelia-modifying agents, we establish a first-of-its-kind high-throughput phenotypic organoid screen. We demonstrate the ability to screen thousands of samples and uncover biological targets and associated small molecule inhibitors which translate to in vivo. This approach is enabled by employing a functional, cell-type specific, scalable assay on an organoid model designed to represent the physiological cues of in vivo Paneth cell differentiation from adult intestinal stem cells. Further, we miniaturize and adapt the organoid culture system to enable automated plating and screening, thereby providing the ability to test thousands of samples. Strikingly, in our screen we identify inhibitors of the nuclear exporter Xpo1 modulate stem cell fate commitment by inducing a pan-epithelial stress response combined with an interruption of mitogen signaling in cycling intestinal progenitors, thereby significantly increasing the abundance of Paneth cells independent of known WNT and Notch differentiation cues. We extend our observation in vivo, demonstrating that oral administration of Xpo1 inhibitor KPT-330 at doses 1,000-fold lower than conventionally used in hematologic malignancies increases Paneth cell abundance. In total, we provide a framework to identify novel biological cues and therapeutic leads to rebalance intestinal stem cell differentiation and modulate epithelial tissue composition via high-throughput phenotypic screening in rationally-designed organoid model of differentiation.

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