Ownership of Human Tissues and Cells
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Use of Cell Culture in Virology for Developing Countries in the South-East Asia Region © World Health Organization 2017
USE OF CELL C USE OF CELL U LT U RE IN VIROLOGY FOR DE RE IN VIROLOGY V ELOPING C O U NTRIES IN THE NTRIES IN S O U TH- E AST USE OF CELL CULTURE A SIA IN VIROLOGY FOR R EGION ISBN: 978-92-9022-600-0 DEVELOPING COUNTRIES IN THE SOUTH-EAST ASIA REGION World Health House Indraprastha Estate, Mahatma Gandhi Marg, New Delhi-110002, India Website: www.searo.who.int USE OF CELL CULTURE IN VIROLOGY FOR DEVELOPING COUNTRIES IN THE SOUTH-EAST ASIA REGION © World Health Organization 2017 Some rights reserved. This work is available under the Creative Commons Attribution-NonCommercial- ShareAlike 3.0 IGO licence (CC BY-NC-SA 3.0 IGO; https://creativecommons.org/licenses/by-nc-sa/3.0/igo). Under the terms of this licence, you may copy, redistribute and adapt the work for non-commercial purposes, provided the work is appropriately cited, as indicated below. In any use of this work, there should be no suggestion that WHO endorses any specific organization, products or services. The use of the WHO logo is not permitted. If you adapt the work, then you must license your work under the same or equivalent Creative Commons licence. If you create a translation of this work, you should add the following disclaimer along with the suggested citation: “This translation was not created by the World Health Organization (WHO). WHO is not responsible for the content or accuracy of this translation. The original English edition shall be the binding and authentic edition.” Any mediation relating to disputes arising under the licence shall be conducted in accordance with the mediation rules of the World Intellectual Property Organization. -
Hybridoma Technology History of Hybridoma Technology
Hybridoma Technology History of Hybridoma Technology What is hybridoma technology? Hybridoma technology is a well-established method to produce monoclonal antibodies (mAbs) specific to antigens of interest. Hybridoma cell lines are formed via fusion between a short-lived antibody-producing B cell and an immortal myeloma cell. Each hybridoma constitutively expresses a large amount of one specific mAb, and favored hybridoma cell lines can be cryopreserved for long-lasting mAb production. As a result, researchers usually prefer generating hybridomas over other mAb production methods in order to maintain a convenient, never-ending supply of important mAbs. Fig 1. Hybridoma technology Inventor: Georges Kohler and Cesar Milstein Hybridoma technology was discovered in 1975 by two scientists, Georges Kohler and Cesar Milstein. They wanted to create immortal hybrid cells by fusing normal B cells from immunized mice with their myeloma cells. For incidental reasons, they had all the requirements fulfilled and it worked in the first attempt. By cloning individual hybrid cells, they established the first hybridoma cell lines which can produce single type of antibody specific to the specific antigen. Their discovery is considered one of the greatest breakthroughs in the field of biotechnology. For the past decades, hybridomas have fueled the discovery and production of antibodies for a multitude of applications. By utilizing hybridoma technology, Sino Biological provides cost-effective mouse monoclonal antibody service, and we can deliver you purified antibodies in 60 days. Steps Involved in Hybridoma Technology Hybridoma technology is composed of several technical procedures, including antigen preparation, animal immunization, cell fusion, hybridoma screening and subcloning, as well as characterization and production of specific antibodies. -
Peptide-Based Scaffolds for the Culture and Maintenance of Primary
www.nature.com/scientificreports OPEN Peptide‑based scafolds for the culture and maintenance of primary human hepatocytes Douglas MacPherson1, Yaron Bram1, Jiwoon Park1 & Robert E. Schwartz1,2* We report here the use of a nanofbrous hydrogel as a 3D scafold for the culture and maintenance of functional primary human hepatocytes. The system is based on the cooperative assembly of a fber‑forming peptide component, fuorenylmethyloxycarbonyl‑diphenylalanine (Fmoc‑FF), and the integrin‑binding functional peptide ligand, Fmoc‑arginine‑glycine‑aspartic acid (Fmoc‑RGD) into a nanofbrous gel at physiological pH. This Fmoc‑FF/RGD hydrogel was formulated to provide a biomimetic microenvironment with some critical features such as mechanical properties and nanofber morphology, which were optimized to support hepatocyte culture. The material was shown to support maintenance and function of encapsulated primary human hepatocytes as indicated by actin staining, qRT‑PCR, and functional cytochrome P450 assays. The designed gel was shown to outperform Matrigel in cytochrome P450 functional assays. The hydrogel may prove useful for liver development and disease models, as well as providing insights into the design of future implantable scafolds for the regeneration of liver tissue in patients with liver disease. Cellular function is determined in part by micro-environmental cues such as soluble factors, extracellular matrix, and cell–cell interactions 1,2. In vitro culture of epithelial cells is ofen associated with epithelial cell dediferentia- tion (i.e. marked by rapid loss of cell-specifc morphology, phenotype, and function)3. Consequently, there is a need for the development and employment of improved materials which can more closely mimic the in vivo microenvironment and reconstitute the appropriate environmental cues in vitro1–3. -
Nano-Silver Particles Reduce Contaminations in Tissue Culture but Decrease Regeneration Rate and Slows Down Growth and Development of Aldrovanda Vesiculosa Explants
applied sciences Article Nano-Silver Particles Reduce Contaminations in Tissue Culture but Decrease Regeneration Rate and Slows Down Growth and Development of Aldrovanda vesiculosa Explants Marzena Parzymies Subdepartment of Ornamental Plants and Dendrology, Institute of Horticultural Production, Faculty of Horticulture and Landscape Architecture, University of Life Sciences in Lublin, Ul. Gł˛eboka28, 20-612 Lublin, Poland; [email protected] Abstract: Aldrovanda vesiculosa is a carnivorous water plant which is endangered by extinction worldwide. The number of natural stands and populations has decreased; therefore, there is a need for its active protection. The best method would be an in vitro culture. One of the main problems is disinfection of the explants. Therefore, it was decided that we should treat the explants with nano- silver particles. The explants were shoot fragments which were disinfected with sodium hypochlorite and then placed in a liquid 1/5 MS medium, supplemented with silver nanoparticles (AgNPs) at a concentration of 5 mg·dm−3. It was observed that AgNPs reduced the number of contaminations but also led to necrosis of the shoots. The shoots, which undertook regeneration in presence of AgNPs, were smaller and did not form traps; however, after being moved to fresh media twice, they started to develop normal leaves. Taking into consideration both disinfection and regeneration rates, it might be advisable to disinfect aldrovanda shoots in sodium hypochlorite only, without AgNPs. The results Citation: Parzymies, M. Nano-Silver of the research might indicate a toxic activity of AgNPs towards water plants, which seems a big Particles Reduce Contaminations in problem, as nanoparticles are commonly used in all the fields of life. -
Hybridoma Technology
HYBRIDOMA TECHNOLOGY Subject : Dr. KRISHNA KUMAR V M.sc,Ph.D,PGDCS Govt. Degree College ,NAIDUPET Email. Id : [email protected] HYBRIDOMA TECHNOLOGY LEARNING OBJECTIVES • TO UNDERSTAND THE NATURE OF ANTIBODY • DISTINGUISH BETWEEN POLYCLONAL AND MONOCLONAL ANTIBODIES • TO LEARN THE TECHNIQUE INVOLVED IN HYBRIDOMA TECHNOLOGY • STEPS INVOLVED IN I) CELL- FUSION III) ANTIBODY PRODUCTION INTRODUCTION ANTIBODY : • Antibodies are specialised proteins that are recruited by the immune system in response to an antigenic stimulus. • Antibodies identify and recognise the foreign antigens and remove them from the body. • Antibodies are produced B lymphocytes (or) B-cells • The production of antibodies is the main function of the humoral response (or) adaptive Immune system TYPES OF ANTIBODIES POLYCLONAL ANTIBODIES: • Antibodies derived from different B lymphocyte cell lines. • Heterogeneous in nature collected from multiple B cell clones. • Recognise and bind to many different epitopes of single antigen. • Primary role is to detect unknown antigens. Monoclonal antibody MONOCLONAL ANTIBODIES (Mabs) : • Antibodies are identical as they are produced by one type of immune cell (or) Clones of single parent cell. • Homogeneous in nature with affinity towards a specific antigen MONOCLONAL ANTIBODIES (Mabs) • Monoclonal antibodies possess two important characteristics, namely high specificity and high reproducibility. • widely used in diagnostic, clinical and therapeutic applications. • Plays a key role in immunotherapy HYBRIDOMA TECHNOLOGY MONOCLONAL ANTIBODIES : • Production of monoclonal antibodies using hybridoma technology was discovered in 1975 by GEORGE KOHLER & CESAR MILSTEIN. • They shared the NOBEL PRIZE in the year,1984. nobelprize.org • In 1990 MILSTEIN produced the first monoclonal antibodies HYBRIDOMA TECHNOLOGY HYBRIDOMA TECHNIQUE : • Well established method to produce monoclonal antibodies specific to desired antigens. -
Producing a Peptide for Use in a Blood Biosensor for Injury
Producing A Peptide For Use In A Blood Biosensor For Injury Detection By Errek Mạnh Trung Phạm Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science in the Biological Sciences Program YOUNGSTOWN STATE UNIVERSITY December 2020 Producing A Peptide For Use In A Blood Biosensor For Injury Detection Errek Mạnh Trung Phạm I hereby release this thesis to the public. I understand that this thesis will be made available from the OhioLINK ETD Center and the Maag Library Circulation Desk for public access. I also authorize the University or other individuals to make copies of this thesis as needed for scholarly research. Signature: ____________________________________________________________ Errek M.T. Phạm, Master’s Student Date Approvals: ____________________________________________________________ Dr. Diana L. Fagan, Thesis Advisor Date ____________________________________________________________ Dr. Jonathan J. Caguiat, Committee Member Date ____________________________________________________________ Dr. David K. Asch, Committee Member Date ____________________________________________________________ Dr. Salvatore A. Sanders, Dean of Graduate Studies Date ii Abstract Conventional hybridoma technology has been used for the selection and production of proteins for biosensor development. However, hybridoma technology is typically a slower and more costly procedure than phage display and produces a less durable end- product. Quicker and more efficient production of a small peptide using phage display has been -
Chapter 11: Virology
CHAPTER 11 Virology Ken Peters USFWS – Bozeman Fish Health Center Bozeman, Montana NWFHS Laboratory Procedures Manual - Second Edition, June 2004 Chapter 11 - Page 1 I. Introduction Detection of aquatic animal viruses historically has been by growth and isolation on living cell cultures appropriately researched and chosen for the propagation of target viruses and species of host. Viral detection can also include immunological and nucleotide testing procedures. The determination of a testing procedure is a complex decision involving factors of cost, timeliness, sensitivity, specificity, efficiency, and available host tissues and technology. For the purposes of the Wild Fish Health Survey, the USFWS has chosen the use of cell culture for initial screening and corroboration of test results using appropriate nucleotide primers of specific viral pathogens in polymerase chain reaction (PCR) tests. Other corroborative tests may also be utilized, including serum neutralization, indirect fluorescent antibody techniques, biotinylated DNA probes, and immuno-dot blot tests (see Chapter 12 - Corroborative Testing of Viral Isolates). The following sections describe the procedures and methods for virology using standard cell culture techniques. Definitions: Several terms are used routinely in virology and throughout this section. A full Glossary of terms can be found in Appendix A. Media Formulations: See Appendix B: Media Used in Tissue Culture and Virology. II. Selection of Appropriate Cell Lines All viral testing will utilize cell lines traceable to cell lines from the American Type Culture Collection (ATCC) when available. At the minimum, cell lines will be tested annually for viral sensitivity and mycoplasma infection: see section VI. Quality Control in Tissue Culture, in Chapter 10 -Tissue Culture of Fish Cell Lines. -
Introduction to Mammalian Cell Culture
Workshop Training Series Biomedical and Obesity Research Core Nebraska Center for the Prevention of Obesity Diseases through Dietary Molecules Introduction to Mammalian Cell Culture April 9, 2019 Yongjun Wang Ph.D. Director of Biomedical and Obesity Research Core Nebraska Center for the Prevention of Obesity Diseases through Dietary Molecules What Is Cell Culture Cell culture is the process by which cells are grown in controlled conditions outside of their native environment. Timeline: key milestone in cell cultures History of Cell Culture http://dx.doi.org/10.5772/66905 Primary vs Cell line Primary cells Cell lines Lifespan and division capacity Limited Indefinite Isolated in the lab or bought from Source Bought from commercial provider commercial provider Care and maintenance Complex and difficult Easy to maintain or proliferate Chromosomal aberration Minimal Several Retention of functional markers and Yes Not always signaling pathways Functional study, diagnosis, Drug development, vaccine and protein Application Gene therapy, et al. production, et al. Three Types of Cells Epithelial-like cells Fibroblast-like cells Lymphoblast-like cells Cell differentiation 3T3L1 cells C212 cells Cell Culture Vessels • Most adherent cells require attachment to proliferate • Polystyrene are treated to become hydrophilic and negatively charged once medium is added • Coating with basic synthetic polymers • Poly-L-lysine • Coating with matrix proteins • Collagen, laminin, gelatin, fibronectin Class II Biological Safety Cabinet The Class II Biological Safety -
Hybridoma Technology & Monoclonal Antibody Production Definition
Sanjib Kr Das B.Sc Hons. In ZOOLOGY 06.06.2020 Assistant Professor (WBES) SEM-IV Dept. of Zoology Paper- CC-10 Immunology Jhargram Raj College UNIT-4 Immunoglobulin Hybridoma Technology & Monoclonal Antibody Production Definition: Hybridoma A clone of hybrid cells formed by fusion of normal lymphocytes with myeloma cells. It retains the properties of the normal cell to produce antibodies or T-cell receptors but exhibits the immortal growth characteristic of myeloma cells. Hybridomas are used to produce monoclonal antibody (mAB). Monoclonal antibody Homogeneous preparation of antibody molecules, produced by a single clone of B lineage cells, often a hybridoma, all of which have the same antigenic specificity. (The term monoclonal refers to the fact that all of the cells in a given hybridoma culture are derived from the single clone of cells, and therefore carry the same DNA). A hybridoma is a fusion product of two cells. B-cell hybridomas are generated by artificially fusing antibody-producing, short-lived lymphocytes with long-lived tumor cells in order to generate long-lived daughter cells secreting large amounts of monoclonal antibodies. In 1975, Georges Köhler and Cesar Milstein figured out how to generate large quantities of antibodies derived from a single B-cell clone. By fusing a normal, activated, antibody-producing B cell with a myeloma cell (a cancerous plasma cell), they were able to generate a hybridoma that possessed the immortal growth properties of the myeloma cell parent and secreted the unique antibody produced by the B-cell parent. Over time, myeloma-cell partners were generated that had lost the ability to synthesize their own immunoglobulin, thus ensuring that the only antibodies secreted into the culture medium were those from the B-cell fusion partner. -
Basic Pluripotent Stem Cell Culture Protocols Maria Borowski∗, Maria Giovino-Doherty, Lan Ji, Meng-Jiao Shi, Kelly P
Basic pluripotent stem cell culture protocols Maria Borowski∗, Maria Giovino-Doherty, Lan Ji, Meng-Jiao Shi, Kelly P. Smith and Joseph Laning, Massachusetts Stem Cell Bank, University of Massachusetts Medical School, Shrewsbury, MA 01545 USA Abstract Stem cell research is a rapidly expanding field with the potential to develop therapeutic agents to treat diseases as well as study disease development from early stages. The culture of human pluripotent stem cells shares many of the same protocols as standard mammalian cell culture. However, the successful culture and maintenance of human pluripotent stem cells (hPSCs) in an undifferentiated state requires additional consider- ations to ensure that cells maintain their key characteristics of self-renewal and pluripotency. There are several basic techniques needed for the culturing of mammalian cells, including thawing frozen stocks, plating cells in culture vessels, changing media, passaging and cryopreservation. The protocols in this document represent a subset of the standard operating procedures used to maintain and culture stem cells at the Massachusetts Human Stem Cell Bank, and have been thoroughly testing and verified. A Stem cell culture considerations Stem cell research is a rapidly expanding field with the potential to develop therapeutic agents to treat diseases as well as study disease development from early stages. However, to fulfill this promise, researchers need to have access to standardized protocols for the development, maintenance and differentiation of these unique cells. Such “best practices” will allow comparisons of different studies and hasten the refinement of these techniques. Such standardization can be driven by resources such as StemBook and by stem cell banks. -
From 2D Culture to Organ-On-Chip Lab on a Chip
Volume 18 Number 9 07 May 2018 Pages 1267–1390 Lab on a Chip Devices and applications at the micro- and nanoscale rsc.li/loc ISSN 1473-0197 CRITICAL REVIEW Jochen Kieninger et al. Microsensor systems for cell metabolism – from 2D culture to organ-on-chip Lab on a Chip View Article Online CRITICAL REVIEW View Journal | View Issue Microsensor systems for cell metabolism – from Cite this: Lab Chip,2018,18,1274 2D culture to organ-on-chip Jochen Kieninger, * Andreas Weltin, Hubert Flamm and Gerald A. Urban Microsensor systems for cell metabolism are essential tools for investigation and standardization in cell cul- ture. Electrochemical and optical read-out schemes dominate, which enable the marker-free, continuous, online recording of transient effects and deliver information beyond microscopy and end-point tests. There has been much progress in microfluidics and microsensors, but the translation of both into standard cell culture procedures is still limited. Within this critical review, we discuss different cell culture formats ranging from standard culture vessels to dedicated microfluidic platforms. Key aspects are the appropriate supply of cells, mass transport of metabolites to the sensors and generation of stimuli. Microfluidics enable the transition from static to dynamic conditions in culture and measurement. We illustrate the parameters oxy- gen (respiration), pH (acidification), glucose and lactate (energy metabolism) as well as short-lived reactive Creative Commons Attribution 3.0 Unported Licence. species (ROS/RNS) from the perspective of microsensor integration in 2D and 3D cell culture. We discuss different sensor principles and types, along with their limitations, microfabrication technologies and mate- Received 1st September 2017, rials. -
Mass Propagation of Plant Cells – an Emerging Technology Platform for Sustainable Production of Biopharmaceuticals
mac har olo P gy : Georgiev, Biochem Pharmacol (Los Angel) 2015, 4:5 & O y r p t e s i n DOI: 10.4172/2167-0501.1000e180 A m c e c h e c s Open Access o i s Biochemistry & Pharmacology: B ISSN: 2167-0501 Editorial Open Access Mass Propagation of Plant Cells – An Emerging Technology Platform for Sustainable Production of Biopharmaceuticals Vasil Georgiev* Center for Viticulture and Small Fruit Research, College of Agriculture and Food Sciences, Florida A & M University, 6505 Mahan Drive, Tallahassee, FL 32317, USA Editorial continuous supply of high quality biomass from exotic, rare, protected, or endangered plants, or plants growing in remote, barely accessible Plants have been used as a source of natural compounds with areas [15]. Recently, several PMIs obtained by PCCT have been released, unique chemical structures and wide range of biological activities since including anti-ageing, calming, and protecting cosmetic supplement time immemorial, and to this day, 11% of the essential drugs for human “ResistemTM” Sederma (www.sederma.fr); UV-protective additive application originate in plants [1]. The expanded demand for medicinal PhytoCellTec™ Solar Vitis, anti-aging and delaying the senescence of plants raise issues of concern about sustainability, conservation and hair follicles supplement PhytoCellTec™ Malus Domestica, and anti- the preservation of natural habitats. Overharvesting of some species aging and UV-protective supplement PhytoCellTec™ nunatak® from for commercial uses reduced the stocks of wild populations and has cell culture of rare and protected plant species Saponaria pumila placed some species under threat [2]. Moreover, most of the medicinal (recognized as an Eco breakthrough at the UN Conference Rio+20) by plants are rare or endemic species, growing under specific climate, Mibelle Biochemistry (www.mibellebiochemistry.com).