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
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
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
Role of serum and supplements in culture medium k.skailash saini
ROLE OF SERUM AND SUPPLEMENTS IN CULTURE MEDIA
Serum is a complex mix of albumins, growth factors and growth inhibitors.
Serum is one of the most important components of cell culture media and serves as a source for amino acids, proteins, vitamins (particularly fat-soluble vitamins such as A, D, E, and K), carbohydrates, lipids, hormones, growth factors, minerals, and trace elements.
Serum from fetal and calf bovine sources are commonly used to support the growth of cells in culture.
Fetal serum is a rich source of growth factors and is appropriate for cell cloning and for the growth of fastidious cells.
Calf serum is used in contact-inhibition studies because of its lower growth-promoting properties.
Normal growth media often contain 2-10% of serum.
Supplementation of media with serum serves the following functions :
Serum provides the basic nutrients (both in the solution as well as bound to the proteins) for cells.
Serum provides several growth factors and hormones involved in growth promotion and specialized cell function.
It provides several binding proteins like albumin, transferrin, which can carry other molecules into the cell. For example: albumin carries lipids, vitamins, hormones, etc. into cells.
It also supplies proteins, like fibronectin, which promote the attachment of cells to the substrate. It also provides spreading factors that help the cells to spread out before they begin to divide.
It provides protease inhibitors which protect cells from proteolysis.
It also provides minerals, like Na+, K+, Zn2+, Fe2+, etc.
It increases the viscosity of the medium and thus, protects cells from mechanical damages during agitation of suspension cultures.
It also acts a buffer.
Due to the presence of both growth factors and inhibitors, the role of serum in cell culture is very complex.
Unfortunately, in addition to serving various functions, the use of serum in tissue culture applications has several drawbacks .
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
TYPES OF SYNCHRONIZATION
(I)PHYSICAL CELL SEPARATION
(II)BLOCKADE
PHYSICAL Vs BLOCKADE SYNCHRONIZATION
CONCLUSION
REFFERENCE
Introduction
History
Scale up in suspension:Stirred culture,Continuous flow culture,Air- lift culture,Nasa bioreactor
Scale up in monolayer culture: Roller bottle culture , multisurface culture,fixed -bed culture
Other type of culture for scaling up: HARV Vessels,STLV vessels
Monitoring of scale up
Conclusion
References
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
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
Role of serum and supplements in culture medium k.skailash saini
ROLE OF SERUM AND SUPPLEMENTS IN CULTURE MEDIA
Serum is a complex mix of albumins, growth factors and growth inhibitors.
Serum is one of the most important components of cell culture media and serves as a source for amino acids, proteins, vitamins (particularly fat-soluble vitamins such as A, D, E, and K), carbohydrates, lipids, hormones, growth factors, minerals, and trace elements.
Serum from fetal and calf bovine sources are commonly used to support the growth of cells in culture.
Fetal serum is a rich source of growth factors and is appropriate for cell cloning and for the growth of fastidious cells.
Calf serum is used in contact-inhibition studies because of its lower growth-promoting properties.
Normal growth media often contain 2-10% of serum.
Supplementation of media with serum serves the following functions :
Serum provides the basic nutrients (both in the solution as well as bound to the proteins) for cells.
Serum provides several growth factors and hormones involved in growth promotion and specialized cell function.
It provides several binding proteins like albumin, transferrin, which can carry other molecules into the cell. For example: albumin carries lipids, vitamins, hormones, etc. into cells.
It also supplies proteins, like fibronectin, which promote the attachment of cells to the substrate. It also provides spreading factors that help the cells to spread out before they begin to divide.
It provides protease inhibitors which protect cells from proteolysis.
It also provides minerals, like Na+, K+, Zn2+, Fe2+, etc.
It increases the viscosity of the medium and thus, protects cells from mechanical damages during agitation of suspension cultures.
It also acts a buffer.
Due to the presence of both growth factors and inhibitors, the role of serum in cell culture is very complex.
Unfortunately, in addition to serving various functions, the use of serum in tissue culture applications has several drawbacks .
INTRODUCTION
HISTORY
NEED OF SYNCHRONIZATION
TYPES OF SYNCHRONIZATION
(I)PHYSICAL CELL SEPARATION
(II)BLOCKADE
PHYSICAL Vs BLOCKADE SYNCHRONIZATION
CONCLUSION
REFFERENCE
Introduction
History
Scale up in suspension:Stirred culture,Continuous flow culture,Air- lift culture,Nasa bioreactor
Scale up in monolayer culture: Roller bottle culture , multisurface culture,fixed -bed culture
Other type of culture for scaling up: HARV Vessels,STLV vessels
Monitoring of scale up
Conclusion
References
Cell culture based vaccine??
Cell cultures involve growing cells in a culture dish, often with a supportive growth medium. A primary cell culture consists of cells taken directly from living tissue, and may contain multiple types of cells such as fibroblasts, epithelial, and endothelial cells.
In the United States, 10 different vaccines for chicken pox, hepatitis A, polio, rabies, and rubella are cultured on aborted tissue from two fetal cell lines known as WI-38 and MRC-5. These vaccines are chicken pox, hep-A, hep-A, hep-A/hep-B, polio, rabies, rubella, measles/rubella, mumps/rubella, and MMR II (measles/mumps/rubella).
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.
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
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.
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
History
Cell culture techniques
Species cloned
Approaches of cell cloning
Monolayer culture- Dilution cloning
Microtitration plate
Suspension culture- Cloning in agar
Cloning in methocel
Isolation of clone
By clonal rings
By suspension clone
Application of cell cloning
Conclusion
Reference
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.
Cellular coning refers to generation of genetically identical cells from parent cells. This presentation teaches differences between cell coning and molecular cloning and various methods of cell cloning. Sample questions are also provided for your review of concept learned
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
8. Biology and characterization of cultured cellsShailendra shera
Immediate environment and environment of surrounding medium governs the various properties of cell. The in vitro condition markedly affects the cellular property of cultured cells. For e.g. Reduction in Cell–cell and cell-material interaction. Therefore, it is imperative to develop understanding of biology of cells in response to various environmental conditions. Characterization of cells helps to identify the origin, purity and authenticity of cells and cell lines.
Bioreactors for animal cell suspension cultureGrace Felciya
1. Types of culture
2. Techniques of cultivating animal cell
3. suspension culture/ Non anchorage dependent
4. Bioreactor consideration
5. Requirements of Bioreactor
6. Reactors used in cultivation
Bioreactors are essential in tissue
engineering, not only because they provide an
in vitro environment mimicking in vivo conditions
for the growth of tissue substitutes, but also
because they enable systematic studies of the
responses of living tissues to various mechanical
and biochemical cues.
Cell culture based vaccine??
Cell cultures involve growing cells in a culture dish, often with a supportive growth medium. A primary cell culture consists of cells taken directly from living tissue, and may contain multiple types of cells such as fibroblasts, epithelial, and endothelial cells.
In the United States, 10 different vaccines for chicken pox, hepatitis A, polio, rabies, and rubella are cultured on aborted tissue from two fetal cell lines known as WI-38 and MRC-5. These vaccines are chicken pox, hep-A, hep-A, hep-A/hep-B, polio, rabies, rubella, measles/rubella, mumps/rubella, and MMR II (measles/mumps/rubella).
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.
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
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.
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
History
Cell culture techniques
Species cloned
Approaches of cell cloning
Monolayer culture- Dilution cloning
Microtitration plate
Suspension culture- Cloning in agar
Cloning in methocel
Isolation of clone
By clonal rings
By suspension clone
Application of cell cloning
Conclusion
Reference
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.
Cellular coning refers to generation of genetically identical cells from parent cells. This presentation teaches differences between cell coning and molecular cloning and various methods of cell cloning. Sample questions are also provided for your review of concept learned
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
8. Biology and characterization of cultured cellsShailendra shera
Immediate environment and environment of surrounding medium governs the various properties of cell. The in vitro condition markedly affects the cellular property of cultured cells. For e.g. Reduction in Cell–cell and cell-material interaction. Therefore, it is imperative to develop understanding of biology of cells in response to various environmental conditions. Characterization of cells helps to identify the origin, purity and authenticity of cells and cell lines.
Bioreactors for animal cell suspension cultureGrace Felciya
1. Types of culture
2. Techniques of cultivating animal cell
3. suspension culture/ Non anchorage dependent
4. Bioreactor consideration
5. Requirements of Bioreactor
6. Reactors used in cultivation
Bioreactors are essential in tissue
engineering, not only because they provide an
in vitro environment mimicking in vivo conditions
for the growth of tissue substitutes, but also
because they enable systematic studies of the
responses of living tissues to various mechanical
and biochemical cues.
A key bottleneck for mammalian cell culture productivity is the extended duration of the process with inoculum seed train and production culture stretching between 4-6 weeks in duration. Introducing flexibility in scheduling and execution of cell culture manufacturing campaigns with via a reduction in process duration can be a key strategy for maximizing facility utilization and facilitating the progression of multiple therapeutics to clinical trials. In this work, we investigated the initiation of CHO cell culture production runs using seed cultures cryopreserved in large disposable bags.
ATCC cell lines and hybridomas are shipped frozen on dry ice in cryopreservation vials or as growing cultures in flasks at ambient temperature. Upon receipt of frozen cells, it is important to immediately revive them by thawing and removing the DMSO and placing them into culture. If this is not possible, store the cells in liquid nitrogen vapor (below −130°C). Do not store frozen cells at temperatures above −130°C as their viability will decline rapidly
Bioreactors are devices in which biological or biochemical processes develop under a closely monitored and tightly controlled environment. Bioreactors have been used in animal cell culture since the 1980s in order to produce vaccines and other drugs and to culture large cell populations. Bioreactors for use in tissue engineering have progressed from such devices.
A tissue engineering bioreactor can be defined as a device that uses mechanical means to influence biological processes. In tissue engineering, this generally means that bioreactors are used to stimulate cells and encourage them to produce extracellular matrix (ECM). There are numerous types of bioreactor which can be classified by the means they use to stimulate cells.
This presentation consists of topics related to oncogene, proto oncogene, Tumor suppresor gene, Ras gene family and structure and functions of tumor suppressor gene.
Stem cells are the cells which have the capability to differentiate into any cells of the body when provided with right stimulus and environment. This presentation teaches about stem cells, characteristics, types and cultivation of stem cells in artificial environment. Sample practice questions are also provided in the end to review the concept learned from this presentation.
Workplace safety is an important aspect to protect personnel against injury or serious accident.In case of animal cell culture safety takes a front seat due to nature of work i.e. handling of human cells and tissues, viruses with high potential to cause infections to humans and other adventitious micro organisms. This presentation presents various methods of safety to protect lab personnel from infectious biological agents.
Media is one of the important components for in vitro cultivation of animal cells. Every animal cells have specific requirements and media are designed by keeping in mind those requirements. However, the basic components and design principle remains the same. Every cell culture media contain carbon source, nitrogen source, trace elements, pH indicator, antibiotics ( although it is not recommended) for high value cell culture applications. While designing media various aspects are considered such as availability, cost effectiveness, types off cells to be grown and regulatory requirements. Tis slide also contains sample MCQs questions
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 slide explains the various basic aspect of animal cell culture, cell line and cell strain, initiation and maintenance of primary cell culture, characteristic of primary cell culture and their applications. It also contains MCQs for practice.
This presentation presents an overview of definition, equipment, various cell culturing methods,
characterization, and applications of animal cell culture. This presentation also contains MCQs to acquaint reader about types of question asked in various competitive examinations.
We all have good and bad thoughts from time to time and situation to situation. We are bombarded daily with spiraling thoughts(both negative and positive) creating all-consuming feel , making us difficult to manage with associated suffering. Good thoughts are like our Mob Signal (Positive thought) amidst noise(negative thought) in the atmosphere. Negative thoughts like noise outweigh positive thoughts. These thoughts often create unwanted confusion, trouble, stress and frustration in our mind as well as chaos in our physical world. Negative thoughts are also known as “distorted thinking”.
How to Create Map Views in the Odoo 17 ERPCeline George
The map views are useful for providing a geographical representation of data. They allow users to visualize and analyze the data in a more intuitive manner.
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.
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
1. Animal cell science and Technology
10. Scaling of cell culture
Shailendra Singh Shera, Ph.D
2. LT.10. Scaling up of cell culture
Outline
1. Scaling up definition
2. Methods of scaling up
a. Scale-up in suspension
b. Scale-up in monolayer
c. Immobilized cell cultures
d. Microcarrier culture
3.Test your understanding
3. Definition: Scaling up
•Scaling up refers to increase in size of cell culture
•Scale-up involves the development of culture systems in stages from (small scale) laboratory to
(large scale) industry
•The methodology adopted to increase the scale of a culture depends on the proliferation of cells
Methods of Scaling Up of cell culture
a. Scale-up in suspension
b. Scale-up in monolayer
c. Immobilized cell cultures
d. Microcarrier culture
4. Scale-up in suspension is the preferred method as it is simpler. Scale-up of suspension culture
primarily involves an increase in the volume of the culture. Small scale generally means the culture
capacity less than 2 litres volume (or sometimes 5 litres).
Stirred suspension cultures:
It is usually necessary to maintain cell strains in stirred suspension cultures, by agitation (or stirring)
of the medium. The stirring of the culture medium is achieved by a magnet encased in a glass
pendulum or by a large surface area paddle. The stirring is usually done at a speed of 30-100 rpm.
This is sufficient to prevent sedimentation of cells without creating shear forces that would damage
cells.
Static suspension cultures:
Some cells can grow in suspension cultures, without stirring or agitation of the medium, and form
monolayer cells. However, static suspension cultures are unsuitable for scale-up.
Scale up in suspension culture
5. •Scaling up in suspension culture involves following elements:
(i) An increase in the volume of the culture medium
(ii) Agitation is done when the depth exceeds 5mm and above 5-10 cm.
(iii) Sparging with CO2 is done to maintain adequate gas exchange.
(iv) Stirring should be between 30-100rpm as:-
(v) Stirring at higher speed creates shear forces that would damage the cells.
(vi) It should be sufficient enough to prevent cell sedimentation.
(vii) Antifoam or Pluronic F68 is added at conc. 0.01-0.1% when serum conc. is 2%.
(viii) Carboxymethylcellulose(1-2%) is added to inc. the viscosity of the medium.
Reactors used for large scale suspension culture
•Stirred Bioreactors
•Continuous flow reactors
•Air lift fermentors
Stirred Tank Bioreactors : These are glass (smaller vessels) or stainless steel (larger volumes)
vessels. These are closed systems with fixed volumes and are usually agitated with motor-
driven stirrers with considerable variations in design details
6. Continuous-Flow Cultures
•The medium enters from inlet side and the cells along with product exit from outlet side.
•Continuous cycle is maintained.
•It can operate at chemostat ( constant chemical constituent in reactor) or turbidostat mode
(constant cell density) in reactor.
Airlift Fermenters Cultures in such vessels are both aerated and agitated by air (5% CO2 in air)
bubbles introduced at the bottom of vessels. The vessel has an inner draft tube through which the
air bubbles and the aerated medium rise.
7. Monolayer Culture This type of culture is essential for anchorage dependent cells. Scaling up of
such cultures is based on increasing the surface area of the substrate in proportion to the number
of cells and the volume of medium and therefore tends to be more complex than suspension
cultures. The available surface area can be increased by using plates, spirals, ceramics and micro
carriers.
Following strategies are used for scaling up of cell culture as monolayer
a. Roux bottle
b. Plastic film
c. Roller bottle
d. Helicell vessels
e. Multitray unit
f. Bead bed reactor
g. Synthetic hollow fibre cartridge
h. Heterogenous reactors
i. Opticell culture system
8. Roux Bottle It is commonly used in laboratory and is kept stationary so that only a portion of its
internal surface is available for cell anchorage. Each bottle provides Ca. 175- 200 cm2 surface area
for cell attachment and occupies 750-1000 cm3 space.
Roller Bottle This vessel permits a limited scale up as it is rocked or preferably rolled so that its
entire internal surface is available for anchorage. Several modifications of roller bottle further
enhance the available surface,
Roux bottle
Roller bottle
https://www.sciencedirect.com/topics/engineering/roller-bottle
https://www.thomassci.com/Laboratory-Supplies/Culture-Bottles/_/ROUX-CULTURE-BOTTLE
9. Multitray Unit
A standard unit has 10 chambers stacked on each other, which have interconnecting channels; this
enables the various operations to be carried out in one go for all the chambers.
Each chamber has a surface area of 600 cm2.
This polystyrene unit is disposable and gives good results similar to plastic flasks.
NuNc Cell factories
10. By immobilizing the cell in cultures, there stability and specificity increases. Two basic mechanisms
used for immobilization of animal cells are; Immurement culture methods and Entrapment culture
methods
Immurement culture– In this type of culture, cells are encapsulated in a polymeric matrix by
adsorption. Matrix used for immobilization are gelatin, polylysine, alginate and agarose.
Entrapment culture– In this type of culture, the cells are held within an open matrix through which
the medium flows freely. The cells may be entrapped within the porous ceramic walls of the unit or
cells also can be enmeshed in cellulose fibers.
Scale up as Immobilized culture
11. Monolayers can be grown on small spherical carriers or micro-beads (80-300 pm diameter)
referred to as micro-carriers. These systems use 90-300 pm dia particles as substrate for cell
attachment. Initially, Dextran beads (Sephadex A-50) were used by Van Wezel in 1967; these
were not entirely satisfactory due to the unsuitable charge of beads and possibly due to toxic
effects. The micro-carriers are made up of any one the following materials (trade names given in
brackets).
i. Plastic (acrobeads, bioplas).
ii. Glass (bioglass, ventreglas).
iii. Gelatin (ventregel, cytodex-3).
iv. Collagen (biospex, biospheres)
v. Cellulose (DE-52/53).
vi. DEAE Dextran (cytodex I, dormacell).
Scale up as microcarrier culture
12. *Practice questions: MCQ
1. For adherent culture scale up can be done as
a. Monolayer
b. Suspension
c. Immobilized
d. None of the above
2. For transformed cells can be scaled up as
a. Monolayer
b. Suspension
c. Immobilized
d. None of the above
3. What do you understand by microcarrier culture scale up
a. Scaling up in suspension
b. Scaling up as monomalyer
c. Scale up using Dextran or glass bead
d. None of the above
4. NUNC cell factories is used for scale up of
a. Adherent cells
b. Non-adherent cells
c. Transformed cells
d. Lymphoblast cells
*Questions adapted from: Practice and Learn Animal cell Science and Technology: Multiple choice question for learning.
Author: Shailendra Singh Shera . Publisher: Amazon Kindle.
13. Suggested reading
1. Watson, J.D., Gilman, M., Witowski J.and Zoller, M. Recombinant DNA, 2nd ed., Scientific
American Books, 1983
2. Glick, B.R. and Pasternack, J.J. Molecular Biotechnology, 3rd ed., ASM Press, 2003
3. Davis J.M. Basic Cell Culture: A Practical Approach, IRL Press, 1998
4. Freshney R.I. Animal Cell Culture a practical approach, 1987
MCQ Practice questions
1. Practice and Learn Animal cell Science and Technology: Multiple choice question for learning.
Author: Shailendra Singh Shera . Publisher: Amazon Kindle.
Available on: amazon.com