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advantages of animal cell culture
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May 10, 2017
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This is the cell culture obtained straight from the cells of a host tissue. This can occur spontaneously or be brought about intentionally using drugs, radiation or vi­ruses. Reagents may be stored at a temperature of -20°C but if cells are to be pre­served it may be necessary to provide liquid nitrogen or a -70°C freezer. All other equipment, such as automatic pipette tips and bottles (lids loosely attached) are autoclaved at 121°C for 20 min. Advantages and disadvantages of animal cloning. This method is called Enzymatic Dissociation. Generally the media is the mixture of in- Organic salts and other nutrients capable of sustaining cells in culture such as amino ac­ids, fatty acids, sugars, ions, trace elements,vitamins, cofactors, and ions. A change in osmolality can affect cell growth and function. It is hoped that learn­ing how to control the development of these cells may offer new treatment approaches for a wide variety of medical conditions. ANIMAL CELL CULTURE MEDIA COMPOSITION. It is now commercially available in the form of liquid plasma kept in silicon ampoules or lyophilized plasma. This is usually done by removing them as gently as possible from the substrate with enzymes. Cell culture is a great process to culture to animal cells but for this media is an important step in the process. Your email address will not be published. Historical Events in the Development of Cell Culture: Claude Bernard proposed that physiological systems of an organism can be maintained in a living system after the death of an organism. One of the earliest and major uses of cell culture is the replication of vi­ruses in cell cultures (in place of animals) for use in vaccine production. Since its inception, the animal cell culture in the twentieth century is related to its usefulness in research. Second was the development of the techniques, such as the use of trypsin to remove cells from culture vessels, necessary to obtain continuously growing cell lines (such as HeLa cells). These include monoclonal antibodies, insulin, hormones, etc. Controlled physiochemical environment (pH, temperature, osmotic pressure, O2, etc. Sterilizing indicators such as sterile test strip are necessary for each steri­lizing batch to ensure that the machine is ope­rating effectively. The Advantages and Limitations of Cell Culture as a Model of Bone Formation. Ø  Cell culture can be used to produce monoclonal antibodies with hybridoma technology. Sato established the basis for the development of serum-free media from cocktails of hormones and growth factors. Pipettes are often plugged with non-absorbent cotton wool before putting into containers for sterilizing. When the cells in the primary culture vessel have grown and filled up all of the available culture substrate, they must be Sub-cultured to give them room for continued growth. The other forms of biological fluids used are coconut wa­ter, amniotic fluid, pleural fluid, insect haemolymph serum, culture filtrate, aqueous humour, from eyes etc. A double distilled or reverse osmosis water supply is essential for preparation of media, and rinsing glassware. In utero infection leads to virus and antibody contamination of foetal and other serum used in cell culture production. When talking about a subject as controversial as animal cloning and transgenic animals, it’s important to evaluate its advantages and benefits, as well as the possible risks. Followings are the purpose of Gene Therapy: a. Swapping harmful mutant alleles with functional ones by selective reverse mutation. The use of the antibiotics penicillin and streptomycin in culture medium decreased the problem of contami­nation in cell culture. Since both normal cells and cancer cells can be grown in cul­ture, the basic differences between them can be closely studied. This is relevant for quality control. Cell-based assays have become increas­ingly important for the pharmaceutical in­dustry, not just for cytotoxicity testing but also for high throughput screening of com­pounds that may have potential use as drugs. Live animals and embryos are used to study effects of some compounds on embryo development. Ø  The micro-environment in the culture vessel can induce many physical, chemical and physiological changes in the cells. Standards of identity and labeling for foods have been a controversial topic for as long as agricultural societies have been marketing food to consumers. It is a form of cellular agriculture.. Cultured meat is produced using many of the same tissue engineering techniques traditionally used in regenerative medicine. Serum containing media (media con­taining serum). These include vaccines for polio, rabies, chicken pox, hepatitis B and measles. A variety of tissue culture plastic-ware is avail­able, the most common being specially treated polystyrene. Advantages of Animal Cell Culture: a. First, for Explant Cultures, small pieces of tissue are attached to a glass or treated in a plastic cul­ture vessel and bathed in culture medium. These hybrid cells (called Hybridomas) are formed by fusing two different but related cells. Animal cell culture has become an essential laboratory technique for the study of biochemical and physiological processes. As cells reach the different stages of culture, you need to characterize them and identify any morphological and functional changes. Cell cultures provide a good model system for studying; b. In stationary culture vessels such as T-flasks and bottles where, although the cells are not kept agitated, they are unable to attach firmly to the sub­strate. Artificial skin for use in treating burns and ulcers is the first commercially available product. The third is the use of cells as replace­ment tissues and organs. However, glassware such as pipettes should be soaked in a suitable detergent, then passed through a stringent washing procedure with thorough soaking in distilled water prior to drying and sterilizing. d. It is the most expensive component of the culture medium. Some cell lines that are no longer considered normal (frequently designated as Transformed Cells) are frequently able to grow either attached to a substrate or floating free in suspension; they are Anchorage-Independent. Carrel and Baker developed ‘Carrel’ or T-flask as the first specifically designed cell culture vessel. In vitro embryonic stem cell culture test helps to reduce the number of live embryo used and the compounds which are toxic toward developing embryo (Gipson and Sugrue, 1994, De Silva et al., 1996). Advantages and disadvantages of in vitro and mouse ascites methods for producing mAb are highlighted in this section. In modern molecular bio­logy, Gene Therapy is an experimental technique that involves insertion of cloned/altered genes into cells using r-DNA tech­nology to replace defective genes causing genetic abnormalities or to prevent poten­tial disorders. The cells are frozen slowly at 1°C/min to -50°C and then kept either at -196°C immersed in liquid N2 (in sealed glass ampoules) or above the liquid surface in the gas phase (screw top ampoules). These are obtainable in various designs to allow a wide range of volumes to be filtered (e.g., Millipore, Gelman). Deterioration of frozen cells has been observed at -70°C, therefore, -196°C (liquid N2) seems to be necessary. Using cell fusion tech­niques, it is also possible to obtain hybrid cells by fusing cells from two different par­ents. b. The regulation of pH is done using a variety of buffering systems. Glutamine is usually added at a final concentration of 2 mM, however, once added to the medium the glutamine is only stable for about 3 weeks at 4°C. The antibiotics chosen should clearly not to be toxic to the cells in culture and may de­pend on the type of contamination experi­enced in the individual laboratory. Tissue culture ware is always chosen to match the procedure. It should contain chemical constitu­ents which the cells or tissues are in­capable of synthesizing. In magnetically rotated spinner flasks or shaken Erlenmeyer flasks where the cells are kept actively suspended in the medium; 2. If the cells form tumours when they are injected into animals, they are considered to be Neo-plastically Transformed. The first is the large-scale production of viruses for use in vaccine production. Serial passaging maintains the homogeneity of the cell types. Third, using these cell lines, scientists were able to develop stan­dardized, chemically defined culture media that made it far easier to grow cells. It is worthwhile to know the […] Advantage And Disadvantage Of Animal Cell Tissue Culture. They can be pur­chased as sterile, disposable filters, or they may be sterilized by autoclaving in suitable filter holders. Controlled and defined physiological con­ditions, c. Homogeneity of cell types (achieved through serial passages). It is important to remember that the cul­ture conditions play an important role in determining shape and that many cell cul­tures are capable of exhibiting multiple morphologies. The second, more widely used method, speeds up this process by add­ing digesting (proteolytic) enzymes, such as trypsin or collagenase, to the tissue fragments to dissolve the cement holding the cells to­gether. Once a surplus of cells is available, they can be treated with suitable cryoprotective agents, such as dimethylsulfoxide (DMSO) or glycerol, carefully frozen and then stored at cryogenic temperatures (below -130°C) until they are needed. Ø  More importantly, the uses of animals in scientific experiments (research) were significantly reduced with the invention of animal cell culture techniques. Some of the advantages and disadvantages are concerned with animal-welfare issues. In addition, some normal cells, such as those found in the blood, do not normally attach to sub­strates and always grow in suspension. Ø  Maintaining the sterile aseptic condition is the most difficult part of cell culture. The Classification of tissue cultures is based on the origin of the cells; which can roughly be divided into "primary culture" and "culture of established cell lines." Ø  High possibility of cross contamination of different types of cells in culture. Industrial Fermentation Process (Batch & Continuous Fermentation), Difference between Batch and Continuous Fermentation, Applications of Restriction Endonuclease Enzymes in Biotechnology (Short Notes), Enzyme Immobilization Methods and Applications (Biotechnology Lecture Notes), Industrial Fermentation Process (Batch, Fed-batch and Continuous Fermentation). Glass­ware, such as pipettes, conical flasks, beakers (covered with aluminum foil) are sterilized in a hot air oven at 160°C for one hour. A pH below 6.8 inhibits cell growth. Once released, the cell sus­pension can then be subdivided and placed into new culture vessels. There are two basic methods for doing this. Two basic culture systems are used for growing cells. The Natural Media used to Pro­mote Cell Growth Fall in Three Categories: 1. Other lines which become immor­tal can continue to divide indefinitely and are called Continuous cell lines. Eagle studied the nutrient requirements of selected cells in culture and established the first widely used chemically defined medium. The procedure for freezing cells is general for all cells in cul­ture. Earle isolated mouse L fibroblasts which formed clones from single cells. The major advantages of primary cultures are the retention of: In many cases, the metabolism of a primary cell culture has greater similarity to in vivo than that seen with sub-cellular fractions used as an exogenous source for biotrans­formation. The incubation temperature will de­pend on the type of cells being cultivated. The resulting hybridomas can produce large quantities of the desired antibody. History of Animal Cell Culture 2. Se­rum is one of the very important com­ponents of animal cell culture which is the source of various amino acids, hormones, lipids, vitamins, poly- amines, and salts containing ions such as calcium, ferrous, ferric, potassium, etc. Ø  Using molecular techniques, primary cells can be transformed and then it can be sub-cultured for unlimited passages. all type of media composes of many essential components which used by cells for growth proliferation and differentiate during culture… c. Homogeneity of cell types (achieved through serial passages) d. Economical, since … Cost factor is a major limitation. The use of contaminated cells for vaccine production may result in contam … Benefits and risks due to animal serum used in cell culture production It is important to maintain conti­nuity in cells to prevent genetic drift and to guard against loss of the cell line through con­tamination and other disasters. The major advantage of using cell culture for any of the above applications is the consistency and reproducibility of results that can be obtained from using a batch of clonal cells. Ø  The cell culture technique can be used for in vitro cytotoxicity studies to test the possible toxicity of compounds or drugs. Both items are very important for storage of liquid media at 4°C and for enzymes (e.g., trypsin) and some media components (e.g., glutamine and serum) at -20°C. One limitation of primary cultures is the ne­cessity to isolate cells for each experiment. b. This is called a Primary Culture. For example, primary cultures of rat myocar­dial cells consisting of synchronously beat­ing cells can be prepared. However, the tissue culture process is not always profitable and there are a plenty of difficulties in maintaining the cells in in vitro conditions. Ø  With the invention of live cell imaging technique and fluorescent tagging methods, many physiological and molecular events in the cells can be visualized in a relatively inexpensive way through the use of in vitro cultured cells. Plasma can also be prepared in the laboratory taking out blood from male fowl and adding heparin to prevent blood coagulation. A wide range of suitable preparations are available from relatively specific antibiotics, e.g., penicil­lin/streptomycin solutions, to broader spectrum antibacterial/antimycotic agents such as kanamycin or amphotericin B. Kohler and Milstein produced the first hybridoma ca­pable of secreting a monoclonal antibody. Human growth hormones produced from recombinant bacteria was accepted for therapeutic use. are also used for culture of animal cells. When using primary cells and limiting the time we study them in culture, we can eliminate the issue of unstable cell lines. Cultured meat is meat produced by in vitro cell culture of animal cells, instead of from slaughtered animals. Suspension Cultures are usually Grown Ei­ther: 1. Different types of synthetic media can be prepared for a variety of cells and tissues to be cultured. Biological fluids such as serum. Lewis and Lewis made the first liquid media consisted of sea water, serum, embryo extract, salts and pep­tones. Ø  Identification of cell type is often difficult since in most of the cases the marker proteins will not express in ample quantity under in vitro conditions. Difference between Batch and Continuous Fermentation, too good Web site for biotechnology student, Your email address will not be published. Abercrombie observed contract inhibition: motility of diploid cells in monolayer culture ceases when con­tact is made with adjacent cells. The medium supplies all the nutrients, growth factors, and required gases for the growth of the cells. [pic][pic][pic][pic]A few of the advantages perceived in the use of biotechnology in agriculture are rapid multiplication of plant species, breeding of resistant varieties, diagnosis and control of disease in crop and livestock, utilization of crop residues and animal waste. Originally, these cell culture tests were done in 96 well plates, but increas­ing use is now being made of 384 and 1536 well plates. Insect cells are widely used as miniature cells factories to express substantial quan­tities of proteins that they manufacture after being infected with genetically engi­neered baculoviruses. This may result in the loss or damage of specific membrane receptors, damage to the integrity of the membrane, and loss of cellular products. All commercial media are formulated in such a way that their final osmolality is around 300 mOsm. Ø  The concentration (C) and time (T) dependent (C X T) effects of compounds such as pharmacologically active molecules, drugs or toxins can be easily studied by cell culture methods. Procedures to isolate cells require the disrup­tion of the tissue, often with proteolytic enzymes. The optimum pH is essential to maintain the proper ion balance, optimal functioning of cellular enzymes and bind­ing of hormones and growth factors to cell surface receptors in the cell cultures. The sec­ond advantage of primary cultures is the retention of tissue specific functions. Please click on the Download Link / Button below to Save the post as a Single PDF file. Harrison cultivated frog nerve cells in a lymph clot held by the ‘hanging drop’ method and observed the growth of nerve fibres in vitro for several weeks. Enter your e-mail address. Littlefield introduced the HAT medium for cell selec­tion. A technique which revolutionizes cell culture by allowing the continuous production and collection of cells, has been developed by scientists at Newcastle University. Advantages of Animal Cell Culture: Ø Physio-chemical environment in the culture such as pH, temperature, osmolarity and level of dissolved gases can be precisely controlled in the in vitro system. Cultured cells are usu­ally described based on their morphology (shape and appearance) or their functional characteristics. Ø  Most of the molecular pathways that taking place inside a cell was elucidated by the use of cell culture techniques. Such culture comprises mostly heterogeneous cells and most of the cells divide only for a limit… In this article we will discuss about Animal Cell Culture:- 1. The cells dissociated from the parental tissue are grown on a suitable container and the culture thus obtained is called primary cell culture. A po­tential supply of replacement cells and tis­sues may come out of work currently be­ing done with both embryonic and adult stem cells. Disadvantages of Serum in Culture Me­dium are: a. That way, we can maintain a more rational and balanced perspective on such a relevant topic. The tests to ensure this, such as growth curves and time of reaching a confluent monolayer, are similar, to those used to ensure that serum batches are satisfactory. Salt, Glu­cose and Amino acids in the growth media determine the osmolality of the medium. Advantages and Disadvantages of Cell / Tissue Culture. d. Interchanging non-functional gene with normal gene through homologous recombination. Disadvantages 7. Cultured cells are widely used alone or in conjunction with animal tests to study the effects of new drugs, cosmetics and chemicals on survival and growth in a wide variety of cell types. Examples of some media are: minimal es­sential medium (MEM), RPMI 1640 me­dium, CMRL 1066, F12, etc. a. In addition to 'milk' produced by livest… The char­acteristics of cultured cells result from both their origin (liver, heart, etc.) Ø  Cells in the culture can be easily quantified by different cell quantification techniques. These techniques can also be used to produce these new proteins in large quantity in cultured cells for further study. ), b. In addition to buffer­ing the medium, there are other growth requirements including amino acids, the requirement for which may vary with cell culture type. In cell culture the cells are maintained in an artificial environment consists of a suitable culture vessel containing a culture medium. One of the major advantages of cell culture is the ability to manipulate the physico-chemical (i.e., temperature, pH, osmotic pressure, O 2 and CO 2 tension) and the physiological environment (i.e., hormone and nutrient concentrations) in which the cells propagate. Need of expertise and technical skill for the development, and regular use of tissue culture. Morphologi­cal or ultra-structural markers can also be examined (e.g., beating heart cells). These cells can then be exam­ined for abnormalities in their chromosomes and genes using karyotyping, chromosome painting and other molecular techniques. Glutamine is also required by most cell lines and it has been suggested that cultured cells use glutamine as an energy and carbon source in preference to glucose, although glucose is present in most defined media. The basis for the animal cell culture media is the balanced salt solution, which are used to create a physiological pH and osmolarity required to maintain the animal cells in vitro or in laboratory conditions. Many cell lines, especially those derived from normal tissues, are considered to be Anchorage-Dependent, that is, they can only grow when attached to a suitable sub­strate. Ham introduced the first serum-free medium which was able to support the growth of some cells. Ø  Chances of chemical and microbial contamination are very high in in vitro methods. These antibodies, called Monoclonal Antibod­ies due to their purity, have many impor­tant clinical, diagnostic, and industrial applications with a yearly value of well over a billion dollars. c. Inserting a new gene into the body to help battle a disease. Addition of serum to the culture media has been an age-old practice. While cul­tured cells can be used to produce many important products, three areas are gen­erating the most interest. Roux maintained embryonic chick cells in a saline culture. Variations in the quality of water used may account for varia­tion in results. Glucose is added as energy source—its concentration varies de­pending on the requirement. Ø  Experience and expertise are required for an effective maintenance most of the cells. In most of the mammalian cell cultures, the temperature is main­tained at 37°C in the incubators as the body temperature of Homo sapiens is 37°C. Advantages and Limitations of Tissue Culture: 1. Advantages of Serum in Culture Medium are: 2. 5. This will insulate the am­poules to slow the freezing process to 1°C/min in a -70°C freezer. In addition, it is possible, by the use of chemicals, viruses and radiation, to convert normal cultured cells to cancer causing cells. The suitability of using PDMS for a microfluidic-based cell-culture system can be summarized as: (1) biocompatible and nontoxic, (2) highly oxygen permeable, and (3) optically transparent, which facilitates microscopic observation. Availability of a wide range of plastic tissue culture reduces the amount of necessary wash­ing up. The term ‘Plant Tissue Culture’ symbolizes the in vitro culture of plant cells. Adherence to good laboratory practice when working with cell cultures is essential for two reasons: first, to reduce the risk of exposure of the worker to any potentially infectious agent(s) in the cell culture, and second, to prevent contamination of the cell culture with microbial or other animal cells (see Aseptic technique and minimization of aerosols). Unless good sterile conditions can be maintained (e.g., using laminar flow hoods) it is ne­cessary to incorporate antibiotics and antimycotics into the media. Jolly observed cell division of salamander leucocytes in vitro. 1. Amniocentesis, a diagnostic technique that enables doctors to remove and culture fetal cells from preg­nant women, has given doctors an impor­tant tool for the early diagnosis of fetal disorders. (adsbygoogle=window.adsbygoogle||[]).push({}). The second is the large-scale production of cells that have been genetically engi­neered to produce proteins that have me­dicinal or commercial value. The Advantages and Disadvantages of Serum - Animal Cell Culture The Advantages and Disadvantages of Serum Serum, as the supernatant from clotted blood of bovine or equine sources has been found to provide high growth-promoting activity for a range of mammalian cell lines. First, there was the development of antibiotics that made it easier to avoid many of the contamination problems that plagued earlier cell culture attempts. Cells that do not at­tach normally to a substrate but re­main in suspension with a spherical shape. In­sect cells will grow best at around 30°C while mammalian cells require a temperature of 37°C. Cultured cancer cells also serve as a test system to determine suitable drugs and methods for selectively destroy­ing types of cancer. Alternatively, ampoules may be frozen in a polystyrene box with 1″ thick walls. When these cul­tures were exposed to tricyclic antidepres­sants that are cardio toxic, beating were observed. 3. Ten times more expensive for same quan­tity of animal tissue; therefore, reasons for its use should be compelling. Serum- free media (media without se­rum). Even though the in vitro culture of prokaryotes and both plant and animal cells are possible now, the term ‘Tissue Culture’ generally denotes to Animal Cell / Tissue culture. These may exhibit characteristics of either parent or both parents. In this case, the cells more closely resemble the cells in the body, genetically and in behavior, and they provide more realistic and applicable data. Carbon dioxide in the medium is in a developed state and the concentra­tion depends on the atmospheric carbon dioxide tension and temperature. Here we take a look at the evolution of cell culture, the rise of 3D, and how the industry’s shift from “static” to “fluidic” is inspiring the next generation of in vitro models. A potential advantage of cell culture technology is that it might permit faster start-up of the vaccine manufacturing process in the event of a pandemic. A simple inverted microscope is essential so that cultures can be examined in flasks and dishes. Primary animal tissue and micro-organisms must not be cultured in or near the cell culture laboratory and the laboratory must be specifically designated for clean cell culture work. Viruses are obligate intracellular parasites that require living cells in order to replicate. Ø  Due to the rapid growth rate of cells artificial culture, there is a high chance of genetic variation within in a cell population. Although animal cell culture was first success­fully undertaken by Ross Harrison in 1907, it was not until the late 1940’s to early 1950’s that several developments occurred that made cell culture widely available as a tool for sci­entists. Please Share the PDF with your Friends, Relatives, Students and Colleagues…, @. primary tissue culture?Several different types of culture are routinely performed. Cells that are attached to a substrate and appear elongated and bipolar, frequently forming swirls in heavy cultures. Fischer developed a chemi­cally defined medium, CMRL 1066. Terms of Service Privacy Policy Contact Us, Gene Therapy: Types, Demonstration and Drawback, Nucleic Acid Hybridization (With Diagram), Microorganisms Associated with Food (Types) | Food Biotechnology, Different Systems or Modes of Microbial Cultures | Microorganism | Biotechnology, Rancidity of Food: Introduction, Types, Factors and Prevention of Rancidity | Food Chemistry | Biotechnology, Classification of Food Starches | Food Chemistry | Biotechnology, Colloidal Systems in Food: Functions, Types and Stability | Food Chemistry. Some cell lines will eventually stop dividing and show signs of aging. Expertise is needed, so that behaviour of cells in culture can be interpreted and regulated. Ø  Cells in culture can be easily characterized by cytological or immune-staining techniques. Thus, the mechanisms that cause the change can be studied. He was considered by some as the father of cell culture. They should be frozen in exponential phase of growth with a suitable preservative, usually dimethylsulfoxide (DMSO). By increasing the atmo­spheric CO2, the pH will be reduced mak­ing the medium acidic. Storage in plastic may result in leaching of toxic substances from the plastic into the wa­ter. Even though cell-culture based meat products are not yet ready for market, the issue of what to call them has already arisen. Enders reported that polio virus could be grown on human embryonic cells in culture. It is sometimes source of contamination by viruses, mycoplasma, prions, etc.. c. It increases the difficulties and cost of downstream processing. Two-dimensional cell cultures have played a role in the drug discovery and development process for decades, however in recent years three-dimensional (3D) cell culture has taken center stage. The culture media is pre­pared in such a way that it provides: a.

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