Scale up means increasing the quantity or volume of cell culture. For animal cells, the scale up strategies are dependent upon cell types or i.e. whether the cells requires matrix for attachment and growth ( adherent cell culture) or grows freely in suspended form in aqueous media. The scaling up principle for adherent cells are just to increase surface area for attachment while for suspension culture is to increase culture volume. This presentation enlightens the reader about different methods of scaling up of cells culture. Readers are also provided with sample questions for better understanding
Scale up means increasing the quantity or volume of cell culture. For animal cells, the scale up strategies are dependent upon cell types or i.e. whether the cells requires matrix for attachment and growth ( adherent cell culture) or grows freely in suspended form in aqueous media. The scaling up principle for adherent cells are just to increase surface area for attachment while for suspension culture is to increase culture volume. This presentation enlightens the reader about different methods of scaling up of cells culture. Readers are also provided with sample questions for better understanding
Primary and established cell line cultureKAUSHAL SAHU
Introduction
Primary Culture
Steps of Primary Culture
Isolation Of Tissue
Dissection And Disaggregation
Types Of Primary Culture
Primary Explants Culture
Enzymatic Disaggregation
Mechanical Disaggregation
Cell Line( Finite & Continuous)
Naming A Cell Line
Choosing A Cell Line
Maintenance Of Cell Line
Conclusion
Reference
PRODUCTION AND MAINTENANCE OF EMBRYONIC STEM CELLSANKUR SHARMA
Embryonic stem cells are pluripotent stem cells and have capacity to differentiate into all type of cells arising from 3 different germ layers i.e., ecto-, meso- and endoderm. In this presentation brief information is given about different methods for production of embryonic stem cells and their maintenance
This presentation is about different staining methods to distinguish between a viable and a non-viable cell. These staining techniques are of immense importance to study different diseased cells such as cancer cells, nerve cells to evaluate the respiratory and metabolic activity of a cell and the potential effects of various drugs to kill the diseased cell.
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
SYNCHRONOUS CULTURES CAN BE OBTAINED IN SEVERAL WAYS:
Physical fractionation .
Chemical appro ach
CENTRIFUGAL ELUTRIATION
Inhibition of DNA synthesis
Nutritional deprivation
SYNCHRONIZATION AT LOW TEMPERATURE
CELLULAR TOTIPOTENCY
SOME HIGHLIGHTS OF CELL SYNCHRONIZATION
REFERENCES
Cell synchronization helps in obtaining distinct sub population of cells representing different stages of cell cycle.It helps in collecting population wide data of cells progressing through various stages of cell cycle. Immortalization, refers to cells having capability of undergoing cell division infinitely. Immortal cells are particularly preferred in cell culture to enable long time storage and use. This presentation teaches about cell synchronization, methods of cell synchronization, cellular transformation, immortalization and mechanism of immortalization.
Biosensors are the analytical device that are used to measure the concentration of analye , these type of biosensors are made with conjugation of enzymes as a biological eliment to quantify a (bio)chemical substance / analyte are reffered to as Enzyme-probe Biosensors .
Biosensors are of many types but focusing on Enzyme biosensors there are 4 main types which are briefly described in this power point presentation .
The MTT assay and the MTS assay are colorimetric assays for measuring the activity of enzymes that reduce MTT or close dyes (XTT, MTS, WSTs) to formazan dyes, giving a purple color The main application allows to assess the viability (cell counting) and the proliferation of cells (cell culture assays)
It can also be used to determine cytotoxicity of potential medicinal agents and toxic materials, since those agents would stimulate or inhibit cell viability and growth
Primary and established cell line cultureKAUSHAL SAHU
Introduction
Primary Culture
Steps of Primary Culture
Isolation Of Tissue
Dissection And Disaggregation
Types Of Primary Culture
Primary Explants Culture
Enzymatic Disaggregation
Mechanical Disaggregation
Cell Line( Finite & Continuous)
Naming A Cell Line
Choosing A Cell Line
Maintenance Of Cell Line
Conclusion
Reference
PRODUCTION AND MAINTENANCE OF EMBRYONIC STEM CELLSANKUR SHARMA
Embryonic stem cells are pluripotent stem cells and have capacity to differentiate into all type of cells arising from 3 different germ layers i.e., ecto-, meso- and endoderm. In this presentation brief information is given about different methods for production of embryonic stem cells and their maintenance
This presentation is about different staining methods to distinguish between a viable and a non-viable cell. These staining techniques are of immense importance to study different diseased cells such as cancer cells, nerve cells to evaluate the respiratory and metabolic activity of a cell and the potential effects of various drugs to kill the diseased cell.
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
SYNCHRONOUS CULTURES CAN BE OBTAINED IN SEVERAL WAYS:
Physical fractionation .
Chemical appro ach
CENTRIFUGAL ELUTRIATION
Inhibition of DNA synthesis
Nutritional deprivation
SYNCHRONIZATION AT LOW TEMPERATURE
CELLULAR TOTIPOTENCY
SOME HIGHLIGHTS OF CELL SYNCHRONIZATION
REFERENCES
Cell synchronization helps in obtaining distinct sub population of cells representing different stages of cell cycle.It helps in collecting population wide data of cells progressing through various stages of cell cycle. Immortalization, refers to cells having capability of undergoing cell division infinitely. Immortal cells are particularly preferred in cell culture to enable long time storage and use. This presentation teaches about cell synchronization, methods of cell synchronization, cellular transformation, immortalization and mechanism of immortalization.
Biosensors are the analytical device that are used to measure the concentration of analye , these type of biosensors are made with conjugation of enzymes as a biological eliment to quantify a (bio)chemical substance / analyte are reffered to as Enzyme-probe Biosensors .
Biosensors are of many types but focusing on Enzyme biosensors there are 4 main types which are briefly described in this power point presentation .
The MTT assay and the MTS assay are colorimetric assays for measuring the activity of enzymes that reduce MTT or close dyes (XTT, MTS, WSTs) to formazan dyes, giving a purple color The main application allows to assess the viability (cell counting) and the proliferation of cells (cell culture assays)
It can also be used to determine cytotoxicity of potential medicinal agents and toxic materials, since those agents would stimulate or inhibit cell viability and growth
Learn about how to create your own cell lines for cytotoxicity assays for use in immunotherapy drug development. Specifically measure target cell death.
On the 31st of July 2014 the Computers 4 Kids head office team took part in the Mozilla Webmaker, Maker Party. This is a slide show of what we got up to.
Principles of cell viability assays by surendra.pptxSurendra Chowdary
1.DYE EXCLUSION ASSAYS:
Dye exclusion assays are the simplest methods that are based on utilization of different dyes such as trypan blue, eosin, congo red, and erythrosine B, which are excluded by the living cells, but not by dead cells.
For these assays, although staining procedure is quite straightforward, experimental procedure may be time-consuming in case of large sample sizes.
a. Trypan blue stain assay:
Trypan blue stain assay has initially been developed in 1975 to measure viable cell count and is still used as a confirmatory test for measuring changes in viable cell number caused by a drug or toxin.
Trypan blue stain, a large negatively charged molecule, is one of the simplest assays that are used to determine the number of viable cells in a cell suspension.
Principle:
The principle of this assay is that living cells have intact cell membranes that exclude the trypan blue stain, whereas dead cells do not.
Cell suspension is mixed with the trypan blue stain and examined visually under light microscopy to determine whether cells include or exclude the stain.
A viable cell will have a clear cytoplasm, whereas a nonviable cell will have a blue cytoplasm.
Reagent preparation:
To perform the trypan blue stain assay, 0.4% trypan blue stain and phosphate- buffered saline (PBS) or serum-free medium are obtained.
Trypan blue stain should be stored in dark and filtered after prolonged storage.
As trypan blue stain binds to serum proteins and causing misleading results, serum-free medium should be used to obtain reliable results.
Assay Protocol:
The cell suspension to be tested is centrifuged at 100 g for 5 min.
The supernatant is discarded and the pellet is resuspended in 1-ml PBS solution or serum-free medium.
Then, one portion of this cell suspension is mixed with one portion of trypan blue stain.
The mixture is allowed to stay at room temperature for 3 min. It is important to note that the cells should be counted within 3–5 min of mixing with trypan blue, as longer incubation periods will lead to cell death and hence reduced viability counts.
Following the incubation, a drop of the mixture is applied to a hemocytometer, which is placed on the stage of a binocular microscope.
Viable cells will remain unstained, and nonviable cells will stain, in the hemocytometer and these cells are counted separately.
.
Calculation:
After counting viable and nonviable cells, the total number of viable cells per milliliter of aliquot is determined by multiplying the total number of viable cells by 2, which is the dilution factor for trypan blue.
Similarly, total number of cells per milliliter of aliquot is determined by addition of number of viable and nonviable cells and multiplying it by 2.
Then, the percentage of viable cells is calculated using the following equation.
% Viable cells = Total number of viable cells per milliliter of aliquot × 100.
Total number of cells per milliliter of aliquot
2.COLORIMETRIC ASSAYS:
Colorimetric assays
This presentation details the definition of cell cytotoxicity and cell viability, the difference between the two term and methods of assessment of cells in culture for presence and absence of cytotoxic chemicals or metabolites.
This presentation is about the basics of cytotoxicity and the possible cellular fates a cell goes through and cellular and molecular level alterations. The nanoparticle related cytotoxicity and its emerging impacts in the environment and health and the disease caused by it is also discussed briefly.
In vitro methods of screening of anticancer agentsNikitaSavita
Cancer- It is the leading cause of mortality. Cancer is the disease which is categorized by abnormal cell growth with the dormant to spread to other parts of body.
Comparison of different methods to measure cell viabilitycreativebioarray22
In vitro assessment of cellular viability has become a generic approach in addressing a vast range of biological questions in many areas of biomedical research. Cell viability assays are often used to screen collections of compounds to determine if the test molecules have effects on cell proliferation or show direct cytotoxic effects that eventually lead to cell death. According to the number and type of cells used, choosing a proper assay method to decided whether the expected outcome is valid or not.
In vitro methods for the assessment of general cellular toxicity,
End-points for the assessment of general cellular toxicity
Specialized cells commonly used in toxicology
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
Salas, V. (2024) "John of St. Thomas (Poinsot) on the Science of Sacred Theol...Studia Poinsotiana
I Introduction
II Subalternation and Theology
III Theology and Dogmatic Declarations
IV The Mixed Principles of Theology
V Virtual Revelation: The Unity of Theology
VI Theology as a Natural Science
VII Theology’s Certitude
VIII Conclusion
Notes
Bibliography
All the contents are fully attributable to the author, Doctor Victor Salas. Should you wish to get this text republished, get in touch with the author or the editorial committee of the Studia Poinsotiana. Insofar as possible, we will be happy to broker your contact.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
3. Cytotoxicity:
It is defined as the toxicity caused due to the action of
chemotherapeutic agents to the living cells.
The Cytotoxicity tests are very important in
nanoparticles as they helps in determination of its
proposed biomedical use.
6. The method for determination of cytotoxicity and cell viability
involves dyes like trypan blue, almar blue, neutral red, coomassie
blue etc.
The method causes differentiation of various cell in terms of colors.
The cell is differentiated on basis of cells which uptake color if they
are living or dead.
The drawback of this method is it does not distinguishes between
cells which are dead and which are in the process of dying.
7. Tritium- labeled thymidine uptake: Tritium-thymidine is
involved in the cell nucleus due to the cell growth, and the amount
of the tritium in the nucleus is then measured using a scintillation
counter. Though the Tritium labeled thymidine uptake assay is
sensitive to determine the influence on the DNA polymerization
activity, it requires radioisotope which causes various concerns.
8. MTT Method: MTT is a colorimetric assay and this method is far
superior to the previously mentioned methods as it is easy-to-use,
safe, has a high reproducibility, and is widely used in both cell
viability and cytotoxicity tests. The MTT assay is the best known
method for determining mitochondrial dehydrogenase activities in
the living cells. In the method, MTT is reduced to a purple formazan
by NADH. MTT forms purple needle shaped crystals in the cells
therefore prior to measuring the absorbance, an organic solvent is
required to solubilize the crystals.
9.
10. WST Assay: Dojindo developed highly water-soluble tetrazolium salts
called WSTs. WSTs produce water-soluble formazans and are suitable for cell
proliferation and cytotoxicity assays. WSTs receive two electrons from viable cells
to generate a yellow, orange, or purple formazan dye. WST-8, a highly stable WST,
is often used in kits. WST-8 formazan, and 1-Methoxy PMS have no cytotoxicity in
the cell culture media, additional experiments may be carried out using the same
cells from the previous assay.
11.
12. Dehydrogenase Based Assay: Dehydrogenase-based assays reflect cell
conditions with more sensitivity than the other assays because they depend
on several elements including dehydrogenase, NAD(H), NADP(H), and
mitochondrial activity. The major difference between CCK-8 and the MTT
assay, other than MTT’s toxicity, is the enzymes involved. The CCK-8
assay involves most of the dehydrogenase in a cell. On the other hand,
MTT only involves mitochondrial dehydrogenase. Therefore, the MTT
assay depends on mitochondrial activity, not the cell itself. Additionally,
CCK-8 is far more sensitive than the MTT assay. Since WST-8 formazan is
water soluble, it does not form crystals like MTT. Therefore, after 1-4
hours of incubation with the CCK-8 solution, measurement of O.D. at 450
nm gives the number of viable cells. No extra steps are required.
13. Comparative in vitro cytotoxicity study of silver nanoparticle
on two mammalian cell lines:
Silver is extensively used for its antimicrobial properties. The exact mechanism which
determines its antimicrobial property is unknown but are believed to act by reacting
with thiol group thereby making them unsuitable for microbial growth. It is also
believed to directly act by reacting with cell membrane causing membrane lysis and
thereby cell death.
The assay used for evaluation of cytotoxicity property was MTT assay method. The
absorbance of the cell was measured at 595nm and the process of assay remained the
same.
Colorimetric assay techniques like alamar blue, neutral red and coomassie blue were
also used for conduction of cytotoxic studies.
14. Comparative In Vitro Cytotoxicity Study on Uncoated
Magnetic Nanoparticles: Effects on Cell Viability,Cell
Morphology, and Cellular Uptake
The cytotoxic studies of Magnetic Iron Oxide Nanoparticles(MIONPs) is important in
order to determine its potential biomedical applications. The MIONPs must be
biocompatible. The potential use of magnetic nanoparticles can be in areas of drug
delivery, cell-labeling, magnetofection, diagnostic imagining (MRIs) etc.
The characterization of uncoated MINOPs was done by X-ray diffraction and
Transmission Electron Microscopy ( TEM). The method for cell viability assay for
cytotoxic studies was done using staining techniques using agents like ethidium
homodimer-1.
15. Li, L. Comparative In Vitro Cytotoxicity Study on Uncoated Magnetic
Nanoparticles: Effects on Cell Viability, Cell Morphology, and Cellular Uptake.
Journal of Nano science and Nanotechnology, 12, 1-8.
Dechsakulthorn, F., & Hayes, A. In vitro cytotoxicity assessment of selected
nanoparticles using human skin fibroblasts. Alternatives to Animal Testing and
Experimentation, 397-400.
Qian, Y. (2014). Enhanced cytotoxic activity of cetuximab in EGFR-positive lung
cancer by conjugating with gold nanoparticles. Scientific Reports.
S Ebada, S. (2008). Methods for isolation, purification and structural elucidation of
bioactive secondary metabolites from marine invertebrates. Nature Protocols.