scholarly journals GMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Angela Tait ◽  
Toby Proctor ◽  
Nick J. I. Hamilton ◽  
Martin A. Birchall ◽  
Mark W. Lowdell
Keyword(s):  
Epithelial Cells ◽  
Gold Standard ◽  
Good Manufacturing Practice ◽  
Raw Material ◽  
Stem Cell Population ◽  
Upper Aerodigestive Tract ◽  
Cell Sheets ◽  
Primary Fibroblast ◽  
Serum Supplement ◽  
The Media

AbstractEngineered epithelial cell sheets for clinical replacement of non-functional upper aerodigestive tract mucosa are regulated as medicinal products and should be manufactured to the standards of good manufacturing practice (GMP). The current gold standard for growth of epithelial cells for research utilises growth arrested murine 3T3 J2 feeder layers, which are not available for use as a GMP compliant raw material. Using porcine mucosal tissue, we demonstrate a new method for obtaining and growing non-keratinised squamous epithelial cells and fibroblast cells from a single biopsy, replacing the 3T3 J2 with a growth arrested primary fibroblast feeder layer and using pooled Human Platelet lysate (HPL) as the media serum supplement to replace foetal bovine serum (FBS). The initial isolation of the cells was semi-automated using an Octodissociator and the resultant cell suspension cryopreservation for future use. When compared to the gold standard of 3T3 J2 and FBS containing medium there was no reduction in growth, viability, stem cell population or ability to differentiate to mature epithelial cells. Furthermore, this method was replicated with Human buccal tissue, providing cells of sufficient quality and number to create a tissue engineered sheet.

Model Pemasaran Usaha Mikro Kecil Dan Menengah (Umkm) Berbasis Online Dalam Mendukung Pariwisata

2017 ◽  
Vol 17 (1) ◽  
pp. 66
Author(s):  
Muhammad Tajuddin ◽  
Abdul Manan
Keyword(s):  
Information Technology ◽  
Small And Medium Enterprises ◽  
System Development ◽  
Internet Technology ◽  
Raw Material ◽  
Integrated Marketing ◽  
International Scope ◽  
Data Base Design ◽  
The Media ◽  
Medium Enterprises

West Nusa Tenggara ’NTB’ is a region of the golden triangle stripe of tourism destination located between Komodo island and Tana Toraja. As the industry backing for small and middle scale businesses getting grow up and have got governmental supporting since economical crises hit Indonesia at the end 1997. In fact, many of those who have not yet optimize the information technology (IT) in running their business. The approach to meet the Small and Medium Enterprises (SMEs) with the proper customer or to find the proper raw material seller is through the media.  Development of internet technology is so rapid, it is a necessity of SMEs using it as a business communications medium, because Internet utility for e-commerce complement has long been developed in both national and international scope.  Furthermore, the research elaborates the integrated e-commerce model and SMEs as an information technology-based model of integrated marketing strategy besides, software creation and its implementation on SMEs in Mataram municipality.  E-commerce and SMEs were designed to use System Development Life Cycle (SDLC) concerning the safety sistem, flowcart diagram and data base design aspects.

Leveraging Big Data to Help Each Learner and Accelerate Learning Science

2017 ◽  
Vol 119 (3) ◽  
pp. 1-24 ◽  
Author(s):  
Philip H. Winne
Keyword(s):  
Randomized Controlled Trial ◽  
Big Data ◽  
Gold Standard ◽  
Learning Analytics ◽  
Controlled Trial ◽  
Raw Material ◽  
Learning Science ◽  
Controlled Trials ◽  
Randomized Controlled ◽  
What Works

Background Today's gold standard for identifying what works, the randomized controlled trial, poorly serves each and any individual learner. Elements of my argument provide grounds for proposed remedies in cases where software can log extensive data about operations each learner applies to learn and each bit of information to which a learner applies those operations. Purpose of Study Analyses of such big data can produce learning analytics that provide raw material for self-regulating learners, for instructors to productively adapt instructional designs, and for learning scientists to advance learning science. I describe an example of such a software system, nStudy. Research Design I describe and analyze features of nStudy, including bookmarks, quotes, notes, and note artifacts that can be used to generate trace data. Results By using software like nStudy as they study, learners can partner with instructors and learning scientists in a symbiotic and progressive ecology of authentic experimentation. Conclusion I argue that software technologies like nStudy offer significant value in supporting learners and advancing learning science. A rationale and recommendations for this approach arise from my critique of pseudo-random controlled trials.

Potential autocrine and paracrine mechanisms of recovery from mechanical injury of renal tubular epithelial cells

1998 ◽  
Vol 274 (3) ◽  
pp. F463-F472 ◽  
Author(s):  
Robert J. Anderson ◽  
Carla J. Ray
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Epithelial Cells ◽  
Mechanical Injury ◽  
Renal Tubular Cell ◽  
Thymidine Uptake ◽  
Tubular Epithelial Cells ◽  
Renal Tubular Epithelial Cells ◽  
The Media ◽  
Renal Tubular

The present studies were done to clarify potential pathways of the nephrogenic repair process. Media removed from mechanically injured vascular smooth muscle cells and LLC-PK1 renal tubular epithelial cells significantly stimulated[Formula: see text]thymidine uptake and cell number in quiescent LLC-PK1 cells, demonstrating the existence of potential autocrine and paracrine pathways of nephrogenic repair. The effect of mechanical injury resulting in release of one or more growth factors into culture media was also found in the opossum kidney OK renal tubular cell line. The nonspecific peptide growth factor antagonist suramin inhibited the effect of media from injured LLC-PK1 cells to stimulate[Formula: see text]thymidine uptake in quiescent LLC-PK1 cells. Exposure of quiescent LLC-PK1 cells to six growth factors, including acidic and basic fibroblastic growth factors (aFGF and bFGF), platelet-derived growth factors AA and BB (PDGF-AA and PDGF-BB), endothelin-2, and hepatocyte growth factor, reproduced the biological responses seen when quiescent LLC-PK1 cells were exposed to media from injured cells. Immunoblotting and enzyme-linked immunosorbent assay experiments demonstrated the presence of aFGF, bFGF, and PDGF-BB but not other candidate growth factors in the media from injured LLC-PK1 cells. A neutralizing antibody directed against bFGF attenuated the effect of media from injured cells to stimulate[Formula: see text]thymidine uptake in serum-starved LLC-PK1 cells. These results demonstrate that mechanical injury to renal tubular epithelial cells results in release of aFGF, bFGF, and PDGF-BB into the media and suggests that bFGF may be involved in an autocrine fashion to promote recovery from injury.

Mechanotransduction of stretch-induced prostanoid release by fetal lung epithelial cells

2006 ◽  
Vol 291 (3) ◽  
pp. L487-L495 ◽  
Author(s):  
Ian B. Copland ◽  
Denis Reynaud ◽  
Cecil Pace-Asciak ◽  
Martin Post
Keyword(s):  
Mechanical Ventilation ◽  
Arachidonic Acid ◽  
Epithelial Cells ◽  
Fetal Lung ◽  
Lung Epithelial Cells ◽  
Cyclic Stretch ◽  
Mitogen Activated Protein Kinase ◽  
Lung Epithelial ◽  
The Media ◽  
Cox 2

Mechanical ventilation is the primary supportive treatment for infants and adults suffering from severe respiratory failure. Adverse mechanical ventilation (overdistension of the lung) triggers a proinflammatory response. Along with cytokines, inflammatory mediators such as bioactive lipids are involved in the regulation of the inflammatory response. The arachidonic acid pathway is a key source of bioactive lipid mediators, including prostanoids. Although ventilation has been shown to influence the production of prostanoids in the lung, the mechanotransduction pathways are unknown. Herein, we established that cyclic stretch of fetal lung epithelial cells, but not fibroblasts, can evoke an extremely sensitive, rapid alteration in eicosanoid metabolism through a cyclooxygenase (COX)-2 dependent mechanism. Cyclic stretch significantly increased PGI2, PGF2α, PGD2, PGE2, and thromboxane B2 levels in the media of epithelial cells, but did not alter leukotriene B4 or 12-hydroxyeicosatetraenoic acid levels. Inhibition of COX-2, but not COX-1, attenuated the cyclic stretch-induced PG increase in the media, suggesting that cyclic stretch primarily affected PG synthesis. Substrate (free arachidonic acid) availability for PG generation was increased because of a cyclic stretch-induced activation of cytosolic phospholipase A2 (cPLA2) via an influx of extracellular calcium and phosphorylation by mitogen-activated protein kinase, p44/42MAPK. The data are compatible with cPLA2 and COX-2 being intimately involved in regulating the injury response to adverse mechanical ventilation.

The renal glomerulus

2015 ◽  
Author(s):  
Marlies Elger ◽  
Wilhelm Kriz
Keyword(s):  
Epithelial Cells ◽  
Mesangial Cells ◽  
Juxtaglomerular Apparatus ◽  
Cell Types ◽  
Stem Cell Population ◽  
Glomerular Filtration Barrier ◽  
Crescent Formation ◽  
Filtration Barrier ◽  
Parietal Epithelial Cells ◽  
Bowman's Capsule

The glomerulus performs its functions with three major cell types. Endothelial cells and visceral epithelial cells (podocytes) lie on the inside and outside of the glomerular basement membrane, and together these three structures form the glomerular filtration barrier. Mesangial cells sit in the axial region. Pathologies of all these regions and cell types can be identified. Parietal epithelial cells lining Bowman’s capsule participate in crescent formation, and at the tubular pole some of these cells seem to represent a stem cell population for tubular cells and podocytes. The extraglomerular mesangium and juxtaglomerular apparatus complete the description of the glomerular corpuscle. The structure of these elements, and how they relate to function, are illustrated in detail.

Interleukin-10 protects cultured fetal rat type II epithelial cells from injury induced by mechanical stretch

2008 ◽  
Vol 294 (2) ◽  
pp. L225-L232 ◽  
Author(s):  
Hyeon-Soo Lee ◽  
Yulian Wang ◽  
Benjamin S. Maciejewski ◽  
Kenny Esho ◽  
Christiaan Fulton ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Lung Injury ◽  
Lung Development ◽  
Mechanical Stretch ◽  
Interleukin 10 ◽  
Cyclic Stretch ◽  
Mechanical Forces ◽  
Type Ii ◽  
Fetal Rat ◽  
The Media

Mechanical ventilation plays a central role in the pathogenesis of bronchopulmonary dysplasia. However, the mechanisms by which excessive stretch of fetal or neonatal type II epithelial cells contributes to lung injury are not well defined. In these investigations, isolated embryonic day 19 fetal rat type II epithelial cells were cultured on substrates coated with fibronectin and exposed to 5% or 20% cyclic stretch to simulate mechanical forces during lung development or lung injury, respectively. Twenty percent stretch of fetal type II epithelial cells increased necrosis, apoptosis, and proliferation compared with control, unstretched samples. By ELISA and real-time PCR (qRT-PCR), 20% stretch increased secretion of IL-8 into the media and IL-8 gene expression and inhibited IL-10 release. Interestingly, administration of recombinant IL-10 before 20% stretch did not affect cell lysis but significantly reduced apoptosis and IL-8 release compared with stretched samples without IL-10. Collectively, our studies suggest that IL-10 may play an important role in protection of fetal type II epithelial cells from injury secondary to stretch.

Characterization of biomaterial-free cell sheets cultured from human oral mucosal epithelial cells

2014 ◽  
Vol 11 (3) ◽  
pp. 743-750 ◽  
Author(s):  
Dong Won Hyun ◽  
Yun Hee Kim ◽  
Ah Young Koh ◽  
Hyun Ju Lee ◽  
Won Ryang Wee ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Free Cell ◽  
Cell Sheets

scholarly journals The Effect of The Addition of Promoting Microbes (PROMI) In Making Media of Pleurotus ostreatus

2020 ◽  
Vol 3 (02) ◽  
pp. 68-77
Author(s):  
Karmila ◽  
Ira Taskirawati ◽  
Baharuddin
Keyword(s):  
Raw Materials ◽  
Raw Material ◽  
Oyster Mushroom ◽  
Growth Time ◽  
Harvest Time ◽  
Mycelium Growth ◽  
Growing Media ◽  
Wood Sawdust ◽  
The Media ◽  
Aleurites Moluccana

In making oyster mushroom growing media, one of the stages that must pass is composting of growing media (baglog). At this stage, it takes a week or more to compile the media, so the media is ready to be inoculated by the fungus. This long composting time, economically slow down production in the cultivation of oyster mushrooms. Thus, using an activator to speed up the composting process is often done. During this time, the most widely used activators to shorten composting time is Effective Microorganism (EM4). In this study, another type of activator with the same function as EM4 was tried, namely Promoting Microbes (PROMI). The purpose of this study to determine the effect (dose) compromise on the making of oyster mushroom growing media to mycelium growth time, pinhead growth time; harvest time; the number of the fruiting body, and the weight of fresh mushrooms.  The raw material used candlenut wood sawdust (Aleurites moluccana Willd) in the making of media. The method used in this study is the making of growing media, composting, sterilization, inoculation of the fungus, and maintenance. Growing media consisted of sawn wood sawdust, PROMI, bran, CaSO4, CaCO3, and water. The treatment used is the addition of compromise to the growing media before composting. The doses of PROMI given are as follows: 0 g (P1); 0.25 g (P2); 0.5 g (P3), and 0.75 g (P4), each of which dissolved in 200 ml of water. This study's results indicate that different PROMI when mixing raw materials influences the time of mycelium closure in growing media and the time of harvesting mushroom fruit bodies. For the time of mycelial closing, giving as much as 0.5 g, PROMI has different control effects.  The provision of PROMI as much as 0.5 g and 0.75 g also influences the control in terms of harvest time.

scholarly journals A Arte e a Ciência de dados na era digital

2019 ◽  
Vol 24 (40) ◽  
Author(s):  
Alberto Semeler ◽  
Marcelo Walter
Keyword(s):  
Binary Code ◽  
Raw Material ◽  
Biological Information ◽  
Digital Culture ◽  
The World ◽  
The Media ◽  
Technological Means ◽  
Visualization Techniques ◽  
Digital Or

Gerar dados é uma ação intrínseca da nossa relação cotidiana com o mundo. Nosso contato com os dispositivos tecnológicos digitais está sempre numa lógica binária: transcodificar nossa existência biológica para o código binário gerando um dilúvio de dados. Deste modo, produzimos dados como uma espécie de commodities (matéria prima). Ao produzir dados alimentamos longas cadeias de bases de dados que movimentam nossa cultura digital. Pretende-se analisar como essa profusão de dados pode afetar o pensamento e teorização sobre arte, seja ela digital ou não, bem como, o que fazer com estes dados no plano da arte. Neste sentido, com o uso de métodos computacionais analíticos e quantitativos, oriundos das ciências dos dados, implicam na produção, análise, e teorizações na arte. Nesse contexto, apresentamos como estudo-de-caso uma aplicação que utiliza técnicas de visualização para auxiliar na análise de obras de artistas, temáticas, cores preferidas, bem como de sua trajetória artística.AbstractGenerating data is an intrinsic part of our daily relationship with the world. Our contact with digital technological means is always in a binary logic: transcoding our biological information to the binary code generating a flood of data. This mode, we produce the data as a kind of commodities (raw material). By collecting data they fed the chains of databases that move our digital culture. It is intended to perfect precisely or do not need to know what to do, whether it is digital or not, and what to do with these data in the art plane. In this sense, with the use of analytical and quantitative computational methods, that is, the data of the media, implies in the production, analysis, and theorizations in the art. In this context, we present as a case study an application that uses visualization techniques to aid in the analysis of works by artists, themes, preferred colors, as well as their artistic trajectory

scholarly journals Keratin intermediate filaments: intermediaries of epithelial cell migration

2019 ◽  
Vol 63 (5) ◽  
pp. 521-533 ◽  
Author(s):  
Sungjun Yoon ◽  
Rudolf E. Leube
Keyword(s):  
Cell Migration ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Biomechanical Properties ◽  
Biochemical Properties ◽  
Cell Junctions ◽  
Epithelial Tissue ◽  
Post Translational Modification ◽  
Cell Sheets ◽  
Epithelial Cell Migration

Abstract Migration of epithelial cells is fundamental to multiple developmental processes, epithelial tissue morphogenesis and maintenance, wound healing and metastasis. While migrating epithelial cells utilize the basic acto-myosin based machinery as do other non-epithelial cells, they are distinguished by their copious keratin intermediate filament (KF) cytoskeleton, which comprises differentially expressed members of two large multigene families and presents highly complex patterns of post-translational modification. We will discuss how the unique mechanophysical and biochemical properties conferred by the different keratin isotypes and their modifications serve as finely tunable modulators of epithelial cell migration. We will furthermore argue that KFs together with their associated desmosomal cell–cell junctions and hemidesmosomal cell–extracellular matrix (ECM) adhesions serve as important counterbalances to the contractile acto-myosin apparatus either allowing and optimizing directed cell migration or preventing it. The differential keratin expression in leaders and followers of collectively migrating epithelial cell sheets provides a compelling example of isotype-specific keratin functions. Taken together, we conclude that the expression levels and specific combination of keratins impinge on cell migration by conferring biomechanical properties on any given epithelial cell affecting cytoplasmic viscoelasticity and adhesion to neighboring cells and the ECM.

Sign in / Sign up

Export Citation Format

Share Document