Introduction
Primary Culture
Steps In Primary Culture
Isolation Of Tissue
Dissection And/Or Disaggregation
Types Of Primary Culture
Primary Explant Culture
Enzymatic Disaggregation
Mechanical Disaggregation
Cell Line( Finite & Continuous)
Naming A Cell Line
Choosing A Cell Line
Maintenance Of Cell Line
Conclusion
reference
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
Introduction.
Properties of Stem Cells.
Key Research events.
Embryonic Stem Cell.
Stem cell Cultivation.
Stem cells are central to three processes in an organism.
Research & Clinical Application of stem cell.
Research patents.
Conclusion.
Reference.
Introduction
Primary Culture
Steps In Primary Culture
Isolation Of Tissue
Dissection And/Or Disaggregation
Types Of Primary Culture
Primary Explant Culture
Enzymatic Disaggregation
Mechanical Disaggregation
Cell Line( Finite & Continuous)
Naming A Cell Line
Choosing A Cell Line
Maintenance Of Cell Line
Conclusion
reference
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
Introduction.
Properties of Stem Cells.
Key Research events.
Embryonic Stem Cell.
Stem cell Cultivation.
Stem cells are central to three processes in an organism.
Research & Clinical Application of stem cell.
Research patents.
Conclusion.
Reference.
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
As opposed to common belief, the measurement of growth in cell culture is fairly simple. Most of the tecchniques that are applied for measurement of microbial growth can be applied to cell culture.Of course with some modification. This presentation exactly explains growth measurement techniques with respect to cell culture. At the end you will also find sample multiple choice questions for practice.
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
Introduction
Definition
History
Why are the transgenic animals being produced
Transgenic mice
Mice: as model organism
Methods of creation of transgenic mice
knock-out mice
Application of transgenic mice
Conclusion
References
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
TYPES OF SYNCHRONIZATION
(I)PHYSICAL CELL SEPARATION
(II)BLOCKADE
PHYSICAL Vs BLOCKADE SYNCHRONIZATION
CONCLUSION
REFFERENCE
Introduction
Terminologies
Types of tissue culture
Applications
Culturing
Sub-culturing
Cryopreservation
Detection of contaminants
In vitro transformation of cells
Cell viability
Rules for working in the Lab
Advantages
Limitations
Constituent of animal tissue culture media and their specific applicationKAUSHAL SAHU
INTRODUCTION
HISTORY
PHYSICOCHEMICAL PROPERTIES OF CULTURE MEDIA
pH
CO2, BICARBONATE AND BUFFERING
OXYGEN
TEMPERATURE
OSMOLALITY
BALANCED SALT SOLUTIONS
CONSTITUENTS OF CULTURE MEDIA
AMINO ACIDS
VITAMINS
SALTS
GLUCOSE
OTHER ORGANIC SUPPLEMENTS
ANTIBIOTICS
SERUM
PROTEINS
NUTRIENTS AND METABOLITES
HORMONES AND GROWTH FACTORS
LIPIDS
MINERALS
INHIBITORS
APPLICATIONS OF CULTURE MEDIA
CONCLUSION
REFERENCES
INTRODUCTION
ROLE IN CELL LINE CHARACTERIZATION
CAUSES OF TRANSFORMATION
METHODS OF TRANSFECTION
CHARACTERISTICS OF TRAANSFORMED CELLS
GENETIC INSTABILITY
IMMORTALIZATION
ABRERANT GROWTH CONTROL
TUMORIGENECITY
CHROMOSOMAL ABERATION
APPLICATION
CONCLUSION
REFERENCE
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
As opposed to common belief, the measurement of growth in cell culture is fairly simple. Most of the tecchniques that are applied for measurement of microbial growth can be applied to cell culture.Of course with some modification. This presentation exactly explains growth measurement techniques with respect to cell culture. At the end you will also find sample multiple choice questions for practice.
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
Introduction
Definition
History
Why are the transgenic animals being produced
Transgenic mice
Mice: as model organism
Methods of creation of transgenic mice
knock-out mice
Application of transgenic mice
Conclusion
References
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
TYPES OF SYNCHRONIZATION
(I)PHYSICAL CELL SEPARATION
(II)BLOCKADE
PHYSICAL Vs BLOCKADE SYNCHRONIZATION
CONCLUSION
REFFERENCE
Introduction
Terminologies
Types of tissue culture
Applications
Culturing
Sub-culturing
Cryopreservation
Detection of contaminants
In vitro transformation of cells
Cell viability
Rules for working in the Lab
Advantages
Limitations
Constituent of animal tissue culture media and their specific applicationKAUSHAL SAHU
INTRODUCTION
HISTORY
PHYSICOCHEMICAL PROPERTIES OF CULTURE MEDIA
pH
CO2, BICARBONATE AND BUFFERING
OXYGEN
TEMPERATURE
OSMOLALITY
BALANCED SALT SOLUTIONS
CONSTITUENTS OF CULTURE MEDIA
AMINO ACIDS
VITAMINS
SALTS
GLUCOSE
OTHER ORGANIC SUPPLEMENTS
ANTIBIOTICS
SERUM
PROTEINS
NUTRIENTS AND METABOLITES
HORMONES AND GROWTH FACTORS
LIPIDS
MINERALS
INHIBITORS
APPLICATIONS OF CULTURE MEDIA
CONCLUSION
REFERENCES
INTRODUCTION
ROLE IN CELL LINE CHARACTERIZATION
CAUSES OF TRANSFORMATION
METHODS OF TRANSFECTION
CHARACTERISTICS OF TRAANSFORMED CELLS
GENETIC INSTABILITY
IMMORTALIZATION
ABRERANT GROWTH CONTROL
TUMORIGENECITY
CHROMOSOMAL ABERATION
APPLICATION
CONCLUSION
REFERENCE
A growth medium or culture medium is a liquid or gel designated to support the growth of microorganisms,cells,or small plants.
Culture media generally comprise an appropriate sourcde of energy and compounds which regulate the cell cycle.
A typical culture media is composed of a complement of amino acids,vitamins,inorganic salts,glucose, and serum as a source of growth factors, hormones, and attachment factors.
In addition to nutrients, the medium also helps maintain pH and osmolarity.
Imporatant amino acids, trace elements, growth factor,hormone,transport protein and adhesion factor are added.
Adhesion factor added are main components of intercellular substance such as fibronectin,collagen and laminin.
Primary purpose of introducing SFM is to promote the specific growth of a particular type of cell.
Exapmple of one serum free media: DCMM-1 SFM. High protein serum free medium, designed for hybridoma cell growth and monoclonal antibody production.
Equipments used , types of culture and media, subculturing, secondary culture, finite & continuous cell lines, cryopreservation and applications of cell culture
The chemical compounds produced by plants are collectively referred to as phytochemicals. Biotechnologists have special interest in plant tissue culture for the large scale production of commercially important compounds. These include pharmaceuticals, flavours, fragrances, cosmetics, food additives, feed stocks and antimicrobials.
Most of these products are secondary metabolites— chemical compounds that do not participate in metabolism of plants. Thus, secondary metabolites are not directly needed by plants as they do not perform any physiological function (as is the case with primary metabolites such as amino acids, nucleic acids etc.). Although the native plants are capable of producing the secondary metabolites of commercial interest, tissue culture systems are preferred.
A cell line is a product of immortal cells that are used for biological research.
Cells used for cell lines are immortal, that happens if a cell is cancerous.
The cells can perpetuate division indefinitely which is unlike regular cells which can only divide approximately 50 times.
Human cell lines
MCF-7 breast cancer
HL 60 Leukemia
HEK-293 Human embryonic kidney
HeLa Henrietta lacks
Primate cell lines
Vero African green monkey kidney epithelial cells
Cos-7 African green monkey kidney cells
And others such as CHO from hamster, sf9 & sf21 from insect cells.
In this modern era, many companies are focusing on the research of monoclonal antibodies to treat cancer and other autoimmune diseases. Hence, cell culture plays an important role to produce therapeutic proteins. To make process efficient we should know alpha and gamma about cell line. The growth of cells in the medium is the most important area or checkpoint. The type of cell line is also discussed here.
Ethnobotany and Ethnopharmacology:
Ethnobotany in herbal drug evaluation,
Impact of Ethnobotany in traditional medicine,
New development in herbals,
Bio-prospecting tools for drug discovery,
Role of Ethnopharmacology in drug evaluation,
Reverse Pharmacology.
This is a presentation by Dada Robert in a Your Skill Boost masterclass organised by the Excellence Foundation for South Sudan (EFSS) on Saturday, the 25th and Sunday, the 26th of May 2024.
He discussed the concept of quality improvement, emphasizing its applicability to various aspects of life, including personal, project, and program improvements. He defined quality as doing the right thing at the right time in the right way to achieve the best possible results and discussed the concept of the "gap" between what we know and what we do, and how this gap represents the areas we need to improve. He explained the scientific approach to quality improvement, which involves systematic performance analysis, testing and learning, and implementing change ideas. He also highlighted the importance of client focus and a team approach to quality improvement.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
The Art Pastor's Guide to Sabbath | Steve ThomasonSteve Thomason
What is the purpose of the Sabbath Law in the Torah. It is interesting to compare how the context of the law shifts from Exodus to Deuteronomy. Who gets to rest, and why?
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
How to Split Bills in the Odoo 17 POS ModuleCeline George
Bills have a main role in point of sale procedure. It will help to track sales, handling payments and giving receipts to customers. Bill splitting also has an important role in POS. For example, If some friends come together for dinner and if they want to divide the bill then it is possible by POS bill splitting. This slide will show how to split bills in odoo 17 POS.
2. APPLICATIONS OF TISSUE CULTURE
1. Studies on intracellular activity eg. cell cycle and
differentiation, metabolism
2. Elucidation of intracellular flux eg. hormonal receptors,
signal transduction
3. Studies related to cell to cell interaction eg. cell adhesion
and motility, metabolic cooperation
4. Evaluation of environmental interactions eg. cytotoxicity,
mutagenesis
5. Studies dealing with genetics eg. Genetic analysis,
immortalization, senescence
6. Laboratory production of medical/ pharmaceutical
compounds for wide range of applications eg. Vaccines,
interferon, hormones
2
3. Animal cell culture are useful for the production of
many pharmaceutically/ medically important proteins
which are as follows:
1. Plasminogen
2. Interferon
3. Blood clotting factors
4. Hormones
5. Monoclonal antibodies
6. Erythropoietin
3
4. Major development’s in cell culture
technology
First development was the use of antibiotics which
inhibits the growth of contaminants.
Second was the use of trypsin to remove adherent cells
to subculture further from the culture vessel
Third was the use of chemically defined culture
medium.
4
5. Culture medium definition:
A growth medium or culture
medium is a liquid or gel
designed to support the growth
of cells
5
6. Media Type Examples
Natural media
Biological Fluids
plasma, serum, lymph,
human placental cord serum,
amniotic fluid
Tissue Extracts
Extract of liver, spleen,
tumors, leucocytes and bone
marrow, extract of bovine
embryo and chick embryo
Clots coagulants or plasma clots
Artificial media
Balanced salt
solutions
Eagle’s BS, Hank’s BS,
Basal media MEM DMEM
Complex media RPMI-1640, IMDM
6
Types of Cell culture media
7. Natural Media
Very useful
Lack of knowledge of the exact composition of these
natural media
7
8. Artificial Media
Serum containing media
Serum-free media (defined culture media)
Chemically defined media
Protein-free media
8
9. Basic Components of Culture
Media
Culture media (as a powder or as a liquid) contains:
amino acids
Glucose
Salts
Vitamins
Other nutrients
The requirements for these components
vary among cell lines, and these
differences are partly responsible for the
extensive number of medium
formulations .
9
10. Natural buffering system
HEPES
Phenol red as a pH indicator (yellow or purple)
Inorganic salt
Amino Acids (L-glutamine)
Carbohydrates
Proteins and Peptides (important in serum-free media.
Serum is a rich source of proteins and includes albumin,
transferrin, aprotinin, fetuin, and fibronectin
Fatty Acids and Lipids
Vitamins
Trace Elements
10
14. Cell Line
Morpholog
y
Species Medium Applications
HeLa B Epithelial Human
MEM+ 2mM Glutamine+ 10% FBS +
1% Non Essential Amino Acids
(NEAA)
Tumourigenicity
and virus studies
HL60 Lymphoblast Human
RPMI 1640 + 2mM Glutamine + 10-
20% FBS
Differentiation
studies
3T3 clone
A31
Fibroblast Mouse
DMEM + 2mM Glutamine +5% New
Born Calf Serum (NBCS) + 5% FBS
Tumourigenicity
and virus studies
COS-7 Fibroblast Monkey DMEM+ 2mM Glutamine + 10% FBS
Gene expression
and virus
replication
studies
CHO Epithelial Hamster
Ham′s F12 + 2mM Glutamine + 10%
FBS
Nutritional and
gene expression
studies
HEK 293 Epithelial Human
EMEM (EBSS) + 2mM Glutamine +
1% Non Essential Amino Acids
(NEAA) + 10% FBS
Transformation
studies
HUVEC Endothelial Human
F-12 K + 10% FBS + 100 µg/ml
Heparin
Angiogenesis
studies
Jurkat Lymphoblast Human RPMI-1640 + 10% FBS Signaling studies
14
15. Common media and their applications
Media Tissue or cell line
IMDM
Bone marrow, hematopoietic progenitor cells, human
lymphoblastoid leukemia cell lines
MEM
Chick embryofibroblast, CHO cells, embryonic nerve cells,
alveolar type cells, endothelium, epidermis, fibroblast,
glia, glioma, human tumors, melanoma
DMEM
Mesenchymal stem cell, chondrocyte, fibroblast,
Endothelium, fetal alveolar epithelial type II cells, cervix
epithelium, gastrointestinal cells, mouse neuroblastoma,
porcine cells from thyroid glands, ovarian carcinoma cell
lines, skeleton muscle cells, sertoli cells, Syrian hamster
fibroblast
RPMI-1640
T cells and thymocytes, hematopoietic stem cells, human
tumors, human myeloid leukemia cell lines, human
lymphoblastoid leukemia cell lines, mouse myeloma,
mouse leukemia, mouse erythroleukemia, mouse
hybridoma, rat liver cells
Nutrient
mixture F-10
and F-12
Chick embryo pigmented retina, bone, cartilage, adipose
tissue, embryonic lung cells, skeletal muscle cells 15
16. Media Supplements
Serum in Media
Basic nutrients
Growth factors and hormones
Binding proteins
Promote attachment of cells to the substrate
Protease inhibitors
Provides minerals, like Na+, K+, Zn2+, Fe2+, etc
Protects cells from mechanical damages during agitation of
suspension cultures
Acts a buffer
Antibiotics
16
17. Aseptic conditions
1. Switch on the laminar flow cabinet 20 mts prior to start
working
2. Swab all bottle tops & necks with 70% ethanol
3. If working on the bench use a Bunsen flame
4. Flame all bottle necks & pipette by passing very quickly
through the hottest part of the flame
5. Avoiding placing caps & pipettes down on the bench; practice
holding bottle tops with the little finger
6. Work either left to right or vice versa, so that all material goe
to one side, once finished
17
18. 7. Clean up spills immediately & always leave
the work place neat & tidy
8. Never use the same media bottle for different
cell lines.
9. If caps are dropped or bottles touched
unconditionally touched, replace them with
new ones
10. Necks of glass bottles prefer heat at least for
60 secs at a temperature of 200 C
11. Never use stock of materials during handling
of cells.
18
19. Contaminant’s of cell culture
Cell culture contaminants of two types
Chemical-difficult to detect caused by
endotoxins, plasticizers, metal ions or traces
of disinfectants that are invisible
Biological-cause visible effects on the culture
they are mycoplasma, yeast, bacteria or fungus
or also from cross-contamination of cells from
other cell lines
19
20. Effects of Biological Contamination’s
They competes for nutrients with host cells
Secreted acidic or alkaline by-products ceases the
growth of the host cells
Degraded arginine & purine inhibits the synthesis of
histone and nucleic acid
They also produces H2O2 which is directly toxic to cells
20
21. Detection of contaminants
In general: turbid culture media, change in growth rates,
abnormally high pH, poor attachment, multi-nucleated cells,
graining cellular appearance, vacuolization, inclusion bodies
and cell lysis
Yeast, bacteria & fungi usually shows visible effect on the culture
(changes in medium turbidity or pH)
Mycoplasma detected by direct DNA staining with intercalating
fluorescent substances e.g. Hoechst 33258
Mycoplasma also detected by enzyme immunoassay by specific
antisera or monoclonal abs or by PCR amplification of
mycoplasmal RNA
The best and the oldest way to eliminate contamination is to
discard the infected cell lines directly
21