Group 4 Project - Cell Culture

From CellBiology


Epithelial Cells in Culture

Cell Culture is the process where cells are derived from an organism, such as a plant or animal and is placed within an artificially controlled environment to stimulate growth. The most common methods of cell culture are tissue and organ culture. The availability of appropriate nutrients and conditions will allow cells that are removed from various tissues and organs to continue to develop in vitro, where the cell acts as an independent unit. Cells developed in vitro will continue to divide, increase in size and grow until affected by an external condition, such as nutrient depletion. Cells within culture can either be genetically identical or may show genetic variation.[1] Cell cultures consist of primary cultures, semi continuous cell cultures and continuous cell cultures.

The process of cell culture consists of the isolation of cells, the maintaining of cells within culture, the cross contamination of a cell line, the manipulation of cultured cells, establishing human cell lines and the development of a generation of hybridomas.[2] The technique of cell culture allows for numerous applications, such as the investigation of the biochemistry of cells, the generation of artificial tissues and has made significant impact in the research of virology through the testing of chemical compounds and drugs on specific cell types. Cell culture research has assisted in manufacturing antibodies, vaccines and cell-produced drugs.[3]


Ross Granville Harrison (1870 - 1959)

Cell Culture arose in the 19th century when physiologist Sydney Ringer developed a salt solution that was able to sustain the beating of an animal heart outside the body, however the principle of tissue culture was first evident in 1885 when Ross Granville Harrison and Wilhelm Roux established the method known as tissue culture.

From 1907-1910 Harrison established the methodology of tissue culture, which became a common technique in the mid 1900s. Harrison utilized the process of tissue cell culture to analyze nerve fibres in vitro. Harrison extracted neural tube and lymph fragments from frog embryos and observed the development of the nerve fibres on a glass slide.[4]

Ross Granville Harrison

Alexis Carrel (1873 - 1944)

From 1910-1923 Alexis Carrel successfully developed the technique for organ transplantation. Carrel further refined his technique to develop organs outside the body. Also in 1910, with the assistance of Montrose Burrows, Burrows and Carrel successfully cultured embryonic and adult tissue from a variety of mammals. Carrel was able to establish subcultured cell lines indefinitely through adding extracts from a chick embryo. In 1912 Carrel successfully subcultured a cell line hundreds of times from heart fragments of a chick embryo, even after an initial contamination scare. Arthur Ebling maintained Carrel’s cell line, through Carrels meticulous aseptic techniques that reduced contamination, until 1946 when the culture was terminated. Carrel also experimented with various devices to improve cell culture techniques. In 1923 Carrel established the cell culture flask, called D-flasks. These flasks allowed the cells to be submerged in a larger medium rather than a hanging drop culture and were able to be maintained for a few weeks rather than a few days without subculturing. [5]

Significant Landmarks in Cell Culture

Charles Lindbergh (1902 - 1974)

Throughout the 1930s Charles Lindbergh, with the assistance of his mentor Alexis Carrel, developed devices that improved tissue and organ culture. Lindbergh developed a glass pump chamber that was successfully in almost 1000 experiments to maintain the functioning of an organ. Lindbergh’s pump chamber, also known as an artificial heart, was further refined to lead to the development of the world’s first heart and lung machines and contributed to the first open heart surgery in the 1950s. This glass chamber consisted of PYREX® glass that eliminated contamination issues and was nontoxic. “The perfusion pumps I designed had to be made of a special glass that would not crack under the heat of sterilization or dissolve to the extent ordinary glass would dissolve in perfusing fluids. Such glass was sold under the name Pyrex[6]

Throughout the 1940-1950s, John Franklin Enders, Thomas Huckle Weller & Fredrick Chapman Robbins conducted significant advances in cell culture techniques, which assisted in virology research, where viruses were developed within cell cultures to manufacture vaccines. Bioreactors were created in the 1950s to produce large quantities (holding approximately 20, 000 liters) of antibodies and cell-produced drugs, where cells performed their ideal function within highly controlled and monitored environmental conditions. Today, cell culture is a significant principle for research. In 2006 the US Congress divided $1 billion to various pharmaceutical companies to develop cell cultured influenza vaccines and there have been significant advances through the establishment of various centers that specialize and support cell culture research.[7]


Cell cultures are divided into three general classes which are Primary cell cultures, Semicontinous cell cultures (low-passage cell lines) and Continuous cell cultures (immortalised cell cultures). The differences between these cell cultures arise from where the cells were taken from. Primary cell cultures consist of cells derived directly from the host organism. The semicontinuous cell cultures are obtained from subcultures of a primary culture and usually consist of diploid fibroblasts that undergo a finite number of divisions. Continuous cell cultures are derived from transformed cells that are generally epithelial in origin. They are classified this way because these cultures grow rapidly, are heteroploid (having a chromosome number that is not a simple multiple of the haploid number) and can be subcultured indefinitely, which is unlike the other two cell culture classes.[8]

Harvesting and Isolating Cells

Cells being cultured in a petri dish

The aim of a cell culture is to produce a large quantity of a particular cell type which is of interest as a pure culture to be able to perform further experiments on. In order to culture cells they need to be isolated from their source. This isolation can be done by using enzymes to dissociate all the cells from the source [9] or the cell culture media can be used to select for the intended cell type. In order to isolate a uniform cell type from a tissue with a mixture of cell types, usually the following general procedure is used. The general procedure to isolate cells is:

1. The extracellular matrix needs to be dissociated.

2. Tissue samples are treated with proteolytic enzymes (eg. trypsin and collagenase) to dissociate either cell adhesion proteins or to degrade the matrix if it is digestable.

3. Using agents such as ethylenediaminetetraacetic acid (EDTA) allows binding or chelating to Calcium ions on which cell-cell adhesions depend on.

4. With gentle agitation the single living cells can be separated. [10][11]

The General Procedure Cell Culture

Cells can be separated using different techniques which usually depend on the properties of the cell type to be isolated. The different isolation techniques include:

  • Centrifugation – Separate cells depending on size.[12]
  • Adherence – This is based on the property of the cell type to adhere strongly to glass or plastic from the cells that adhere less strong. To increase the specificity of binding antibodies can be used to specifically bind to the cell type of interest. In this process antibodies specific for the cell type of interest are attached on to matrices (such as collagen, polysaccharide beads or plastic). Then to recover the cells first there is gentle shaking applied and then followed by either treatment with trypsin or by degrading the matrix itself depending on the matrix used.[13][14]
  • Antibody coupled to a fluorescent dye – This technique uses antibodies coupled to a fluorescent dye to attach and label specific cell types. Then all the cells are placed into an electronic fluorescence-activated cell sorter where the cells that are labelled are separated from the unlabled ones.[15]
  • Microdissection – Tissue slices which have been prepared for microscopic examination are used to obtain the cells of interest.[16]

If an enzyme is used to dissociate the cells from the source then the resulting suspension would be spread over a flat surface (eg. bottom of culture flask, or petri dish) in order to culture the cells.

Growth Media For Cell Cultures

All cell cultures need to have growth media. In order to culture cells it is important that they can receive the relevant nutrients, minerals and in some cases hormones or growth factors required for the cells to grow [17]. To be able to culture the preferred cell type it is quite important to pick the correct growth media. This selection of growth media can also help to purify the cell culture. It can be done by using a specific growth media rather than a general one in order to stop other types of cells also growing.

There are many types of different growth media available and are either defined or undefined media. Defined media is growth media in which all the quantities of the ingredients are known, whereas undefined media has some complex ingredients and all the ingredients involved are in unknown amounts. Growth media can be varied in pH, glucose concentration, growth factors and the presence of different nutrients. The factors which can be varied can also be used to help isolate and grow a particular type of cell. Apart from the growth media cells also need the right temperature and appropriate gas mixture to be able to grow. The conditions must be ideal for the particular cell which is being cultured in order for the cell culture to be successful.[18]

A Cell Incubator

A cell incubator is used in order to keep the cells in a desirable state to allow them to proliferate. Most human cells are placed into cell culture incubators with a temperature of 37°C, relative humidity of 95% and 5% carbon dioxide[19].

Important Cell Culture Techniques

There are two different ways to culture cells which are suspension and adherent cultures. Suspension cultures are used for cells which naturally exist without adhering to any surfaces. Adherent cultures are used for cells which require to be attached to a surface. Both types of cell culture would need growth media, however in suspension cultures the growth media would most likely be in a liquid form and in adherent cultures it would be solid to allow the cells to attach and proliferate.[20] The following table demonstrates the differences between the adherent and suspension cultures.

A Table showing the differences between Adherent and Suspension Cultures

Properties Adherent Cultures Suspension Cultures
Cell Types Most cells can be cultured this way Cells which are adapted to suspension cultures or non-adhesive cultures
Passaging Passaging required at certain intervals Passaging is much easier, can dilute culture to stimulate growth
Ability to see cells Allows easy visualisation of cells Harder to view cells
Cell Dissociation Cells dissociated enzymatically or mechanically Not require enzymatic or mechanical dissociation
Cell Growth Surface area limits growth Cell concentration in medium limits growth
Uses Used for cytology, harvesting products continuously and also research applications Used for bulk protein production, batch harvesting and also research applications

Table adapted from [21][22].

Cells need to be cultured using very sterile techniques to avoid cross-contamination of the cell type of interest with other cell types. This is done by using aseptic technique which is a process in which the equipment used to culture the cells are constantly sterilised and the process limits the chance of contamination[23]. There is no guarantee however that this technique is perfect and thus it is recommended that the researchers always verify the cells which they have cultured and make sure it is the cell type of interest. Once verification is complete and the cell of interest has been found it can be subcultured to let that cell type to proliferate.


Human cell lines

Cell culture has beneficial uses to help improve human health. Many different vaccines have been made with the help of this technique which include measles, mumps, rubella, polio and hepatitis A. Therapeutic proteins have also been made using cell cultures of animal cells as hosts. Some of these proteins include Erythropoietin (EPO), Tissue plasminogen activator (TPA) and γ-interferon. The technique of cell culture has also found use assisting the production of cell therapy products and gene therapy products.[24]

Plant tissue culture now has direct commercial applications as well as value in basic research into cell biology, genetics and biochemistry. The techniques include culture of cells, anthers, ovules and embryos on experimental to industrial scales, protoplast isolation and fusion, cell selection and meristem and bud culture. Applications include: cross-pollinate distantly related species and then tissue culture the resulting embryo which would otherwise normally die,for production of doubled monoploid plants from haploid cultures to achieve homozygous lines more rapidly in breeding programmes, usually by treatment with colchicine which causes doubling of the chromosome number, certain techniques such as meristem tip culture can be used to produce clean plant material from virused stock, such as potatoes and many species of soft fruit, micropropagation using meristem and shoot culture to produce large numbers of identical individuals, to cross distantly related species by protoplast fusion and regeneration of the novel hybrid, as a tissue for transformation, followed by either short-term testing of genetic constructs or regeneration of transgenic plants. [25]

A plant breeder may use tissue culture to screen cells rather than plants for advantageous characters, e.g. herbicide resistance/tolerance. Large-scale growth of plant cells in liquid culture inside bioreactors as a source of secondary products, like recombinant proteins used as biopharmaceuticals. Micropropagation is widely used in forestry and in floriculture. Micropropagation can also be used to conserve rare or endangered plant species.Somatic embryogenesis which is the development of embryos from vegetative cells rather than from union of male and female gametes.[26]

Plant Tissue Culture

Some species are unruly to somatic embryogenesis; so are some cultivars within the same species. The recent findings that mutants and epigenetic variants, impaired in somatic embryogenesis, can be rescued by addition to the medium of conditioning factors (glycoprotein) secreted by embryogenic cultures, opens new and exciting perspectives, particularly if these conditioning factors prove to be non specific.

Plant Tissue Culture Lab

The two main applications of plant tissue culture in horticultural production are in propagation and sanitation. Tissue culture is being used effectively in asexual clonal propagation of various horticultural crops. Orchids, carnations, roses, gerbera, chrysanthemums, potatoes, strawberries, apples and coffee have all been successfully cloned in vitro and the list is increasing daily. Recovery of pathogen - free plants by meristem tip culture has become a common practice in producing virus-free stock material of vegetatively propagated plants. Other areas where tissue culture can contribute to horticulture include embryo culture and germplasm preservation and storage.[27]

Importance of Plant Tissue Culture in Biotechnology

Cell suspension culture in liquid medium is a relatively young field of biotechnology. It involves the large scale culture of isolated plant cells under conditions which induce them to synthesize the natural secondary metabolites characteristic of the parent plants from which they were obtained.The techniques of callus culture and cell suspension culture have been reviewed in recent years, in particular from the viewpoint of studying biosynthesis and metabolism of steroids and cardiac glycosides. Biotechnologists are also trying to increase the synthesis of natural compounds or new compounds by higher plant cell culture as a result of mixing or feeding transformable precursors in the culture medium.Biotechnologists are also trying to augment the synthesis of medicinally important alkaloids in culture by means of a fungal elicitor.[28]

Tissue culture growth medium

Biotechnologists are also trying to modify the genetics of cultured cells in three ways:

(i) mutagenesis and selection of cell lines in cell suspension culture, (ii) transplantation of foreign genetic material in protoplasts by means of genetic engineering, and (iii) somatic hybridization by the fusion of distantly related plant protoplasts just to widen the genetic diversity of hybrids.[29].

Application of Organ Culture

Models of growth, differentiation and development of organ basics can be studied and the influence of a range of factors like hormones, vitamins, etc. on these parameters, can be assessed. Action of drugs, carcinogenic agents, etc. on the animal organ is studied in vitro, at least to serve as a guide for the events in whole animals.The most important application of organ culture is the manufacture of tissues for implantation in patients. This is called tissue engineering. Human skin has been successfully produced in vitro and used for transplantation in more than 500 cases of serious burns, ulcers etc. The final purpose of tissue engineering is to reconstitute body parts in vitro for application as grafts or transplants, and as models for studies on drug delivery and action. It is predictable that cartilage tissue developed in vitro (artificial cartilage) will be accessible for human implantation in case of injuries, arthritis, etc. Researches using rabbits have produced promising results. It is hoped that studies will allow the culturing and constitution of bones, liver, pancreas, etc. [30]

The basis of explant is also the patient itself or the prepuce of new born babies. The use of a synthetic polymer PGA, permits the new born skin to grow without scars. This artificial skin is used to cover the wound until the patient's skin is cultured and artificial skin is acquired for gratfing.

The keratinocytes manufacturing up the bulk of the epidermis is trypsioized. These cells are cultured in vessels, the bottom of which is covered with irradiated 3T3 fibroblast cell line. Propagation of keratinocytes is inspired by certain products from fibroblasts. Keratinocytes form colonies, which is again dissociated and cultured. The process is repeated till a nonstop sheet of pure epithelium is made. This sheet is detached, cleaned and used for grafting. Rejection of implant can be evaded by taking the explant from the patient itself. A 1.7 m2 artificial skin can be acquired from a 3 cm2 skin in 34 weeks-a SOOO-foid increase. Artificial skin graft have been used to fix effectively several skin defects, skin ulcers, etc. All essential components of skin will be regenerated in about 5 years, after the grafting of the artificial skin.[31]

In the field of mutagenesis and selection of cell lines in vitro and the exploitation of totipotency, biotechnologists are trying to improve plants e.g. by producing crop species that are more resistant to drought, disease, poor soil conditions, chemical pesticides and herbicides.Another goal of biotechnology using the plant tissue culture technique is to produce plants that would provide their own usable nitrogen. [32]


  • Expertise is needed, so that behaviour of cells in culture can be interpreted and regulated.
  • Ten times more expensive for same quantity of animal tissue; therefore reasons for its use should be compelling.
  • Unstable aneuploid chromosome constitution.[33]

Potential Cell Culture Problems

As with all techniques cell culture has some problems with it as well. The cells being cultured can grow to fill the available area. This can lead to some problems which include:

  • The depletion of nutrients in the growth media
  • Deposition and accumulation of apoptotic/necrotic cells
  • Cell cycle arrest (which can stop dividing known as senescence) can be stimulated by cell to cell contact
  • Cellular differentiation can result from cell to cell contact[34]

There are various ways to avoid these problems by manipulating the cell cultures. To replace nutrients depleted the growth media can be removed and then replaced. To overcome the problem of senescence a subculture can be performed which involves transferring a small number of cells into a new culture dish. Both animal cells and plant cultures can be cultured but the appropriate growth media and conditions must be met. A major difference between plant and animal cell culture is the ability of many of the plant cells, with appropriate signalling and manipulation, to form any of the cell types or tissues of a plant[35]. This feature is known as totipotency.

Limitations of Haploids

  • In many crops, the application of this technique is not yet possible since the technique for haploid production is not available.
  • In many other crops, its application is not feasible because large numbers of haploid plants are not easily obtained.
  • High cost of obtaining haploids and doubled haploids is still a major problem.
  • A nonrandom recovery of haploids may reduce the spectrum of variability recovered.
  • A sophisticated tissue culture laboratory and a dependable greenhouse are essential for success.
  • Specialized skill for carrying out the various operations is required.
  • Occurrence of grametoclonal variation may limit the usefulness of pollen embryos for genetic transformation/gene transfer.
  • High frequency of albinos are produced in anther cultures of monocots, especially cereals.[36]

Limitations of Micropropagation

  • Expense
  • Species specificity
  • Production scheduling
  • Contamination
  • Variability
  • Acclimatization [37]

Limitations of Somatic Embryogenesis

Although somatic embryogenesis in banana is well established with the availability of standard techniques, the initation of an ECS cannot still be considered a routine procedure. This is mainly due to the low embryogenic response of banana tissues, the long time to obtain an embryogenic cell suspension, the risk of somaclonal variation and the occurence of contamination. Main problem with the use of edible bananas is that the embryogenic callus needs to be initiated from differentiated tissues.[38]

Current Research & Advances

Current Success and Future Directions in Ovarian Tissue Culture

Female cancer patients have been offered 'banking' of gametes before starting fertility which can lead to the threatening of cancer therapy. The transplants of fresh and frozen ovarian tissue between healthy fertile and infertile women have established the usefulness of the tissue stored for reinstatement of endocrine and fertility function. Additional methods, like follicle culture and isolated follicle transplantation, are in development.

Experiments on ovarian tissue from non-human primate models and from agreeable fertile and infertile patients benefit from a multidisciplinary approach. The new discipline of oncofertility needs professionalization, multidisciplinarity and mobilization of funding for basic and translational research. [39]

Gene Transcription in the Mammary Gland

Several mammary culture systems have been produced to facilitate the research on the regulation of gene transciption in the mammary gland. The whole organ and explant cultures have been good value in recognising the role of specific hormones in both the growth and differentation of mammary tissue and the induction of milk proten gene expression.

These cultures have a limited lifespan and are not useful for studies at the cellular level. Epithelial cells can be isolated from mammary tissue, maintained in culture and induced to distinguish with lactogenic hormones. The applications of such primary cultures has established the importance of the cellular substratum in the differentiation process. A major disadvantage of this system, in addition to the short lifespan of the cells is the considerable amount of starting material required.[40]

Culturing Cells on a Lab Chip

The technique of cell culture requires an adequate amount of nutrients at all times in order for the cells to be able to grow. Besides this the technique requires certain manual tasks in order to continue a cell culture without inhibiting the growth of the cells. Irena Barbulovic-Nad, Sam H. Au and Aaron R. Wheeler have worked out a method to create cell cultures that will grow on a lab chip. This process uses digital microfluidics to successfully culture cells. This system uses only a fraction of the actual nutrients, hormones and minerals which are normally required and also allows automated subculturing of the cells to avoid problems to do with the area of the cells to proliferate. This technique will be vital in future experiments to efficiently and fairly quickly produce a culture of cells.[41]

The Immune Response

In 2010 the Animal Parasitic Diseases Laboratory conducted a study into the affects of organic extracts upon the immune system and tumor cell activity. The Laboratory utilized in vitro chicken lymphocytes and macrophages to determine the affect of organic extracts, such as milk thistle, turmeric, reishi mushrooms and shitake mushrooms. It was determined that the organic extracts induced cell proliferation and nitric oxide production in the chicken lymphocytes and macrophages, when compared to a controlled untreated in vitro cell. This production and proliferation resulted in the inhibition of tumor cell growth within in vitro chickens. The Laboratory concluded that the medicinal organic plants and mushrooms had a favorable affect upon the immune system of the in vitro chickens.[42]


Aneuploid – an abnormal number of chromosomes

Aseptic – system that prevents infection by being free of microorganisms

Biosynthesis – the formation of chemical compounds, derived from living organisms

Callus Culture - a tissue culture technique that requires growth regulators and extracts from an organ or tissue culture

Centrifugation - an instrument that rotates at high speed to separate substances of different concentrations

Continuous/Immortalised Cell Cultures – cells that are derived from transformed cells that are generally epithelial in origin

Cultivars – a plant variety that is developed and maintained through cultivation

Diploid – a cell with double the number of chromosomes

Epigenetic – the process where genetic information is modified by environmental influences, which is expressed within the behaviour of an organism

Fibroblasts – a connective tissue cell that contributes to forming collagen fibres

Floriculture – cultivation of flowering plants

Germplasm – the genetic resources available for an organism

Glycosides – an organic compound derived from plant to produce a sugar and non-sugar compound

Growth Media – a liquid or gel designed to support the growth of microorganisms or cells

Haploid – a cell with a complete set (half the number of a diploid cell) of chromosomes

Heteroploid – a cell with an abnormal number of chromosomes

In Vitro - where a procedure occurs within a controlled external environment rather than in the living organism

Meristem – the embryonic tissue of plants

Micropropagation – a plant tissue culture technique, where offspring are cloned from the tissue of a single plant

Monocots – type of flowering plant

Mutagenesis – the development and origin of a mutation

Passaging – the process of sub-culturing cells

Primary Cultures – cells that are derived directly from a host’s tissue or organ

Protoplast – the contents of a living cell within a cell membrane

Sanitation – the process and application of measures to ensure the prevention of disease and contamination

Semicontinuous Cell Cultures/Low-Passage Cell Lines – cells that are obtained from subcultures of the cells of primary culture

Senescence – the processing of getting older

Somaclonal – the term used to describe genetic variation that arises within plants due to plant tissue culture

Somatic – affecting the body wall of an organism

Subculture – a cell culture made by transferring cells from a previous culture to a fresh growth medium

Helpful Links

ATCC - The Global Bioresource Center that focuses in Cell Culture

European Collection of Cell Cultures (ECACC) that preserves and distributes cell lines

Genentech- a biotechnology company focusing in human genetic information

Landmarks in Cell Culture

National Cell Culture Center (NCCC) that produces cell lines for fundamental research

PubMed- a datatbase that provides research articles related to Cell Culture

SkinEthic- laboratory focusing in tissue engineering

Tissue Engineering Replacing organs or tissues with lab-created counterparts; engineered kidneys, livers and hearts


  1. Chaudry, Arshard. Cell Culture
  2. Lonza Cell Culture & Process Development
  3. Ryan, John A. Introduction to Animal Cell Culture
  4. Schiff, Judith Ann. (2002) An Unsung Hero of Medical Research
  5. Nobel Alexis Carrel (1967)
  6. Lindbergh, C. A. (1978) Autobiography of Values, Page 136, 1978, Harcourt Brace Jovanovich, New York and London.
  7. Corning Life Sciences - Celebrating a Century of Cell Culture 1907-2007
  8. Willey J.M., Sherwood L.M., & Woolverton C.J. (2008). Prescott, Harley, and Klein's Microbiology (7th ed., pp. 866). New York: McGraw-Hill.
  9. Ryan US. Isolation and culture of pulmonary endothelial cells. Environ Health Perspect. 1984 Jun;56:103-14. Review. PubMed PMID: 6090112; PubMed Central PMCID:PMC1568192.
  10. Alberts B., Johnson A., Lewis J., Raff M., Roberts K. and Walter P. (2002). Molecular Biology of the Cell (4th ed.). New York: Garland Science.
  11. Alberts B., Bray D., Hopkin K., Johnson A., Lewis J., Raff M., Roberts K. and Walter P. (2004). Essential Cell Biology (2nd ed.). New York: Garland Science.
  12. Alberts B., Johnson A., Lewis J., Raff M., Roberts K. and Walter P. (2002). Molecular Biology of the Cell (4th ed.). New York: Garland Science.
  13. Alberts B., Johnson A., Lewis J., Raff M., Roberts K. and Walter P. (2002). Molecular Biology of the Cell (4th ed.). New York: Garland Science.
  14. Alberts B., Bray D., Hopkin K., Johnson A., Lewis J., Raff M., Roberts K. and Walter P. (2004). Essential Cell Biology (2nd ed.). New York: Garland Science.
  15. Alberts B., Bray D., Hopkin K., Johnson A., Lewis J., Raff M., Roberts K. and Walter P. (2004). Essential Cell Biology (2nd ed.). New York: Garland Science.
  16. Alberts B., Johnson A., Lewis J., Raff M., Roberts K. and Walter P. (2002). Molecular Biology of the Cell (4th ed.). New York: Garland Science.
  17. Cooper G. M. & Hausman R. E. (2000). The Cell: A Molecular Approach (2nd ed.). Bethesda, Md: Sinauer Associates.
  18. Madigan M. T., Martinko J. M., Dunlap P. V. & Clark D. P. (2009). Brock Biology of Microorganisms (12th ed.). San Francisco, CA: Pearson Benjamin Cummings.
  19. Triaud F., Clenet D. H., Cariou Y., Neel T. L., Morin D. and Truchaud A. Evaluation of Automated Cell Culture Incubators. Journal of the Association for Laboratory Automation. 2003;8(6):82-86.
  20. Dorfman NA, Civin CI, Wunderlich JR. Susceptibility of adherent versus suspension target cells derived from adherent tissue culture lines to cell-mediated cytotoxicity in rapid 51Cr-release assays. J Immunol Methods. 1980;32(2):127-39. PubMed PMID: 7358991.
  21. Chu L, Robinson DK. Industrial choices for protein production by large-scale cell culture. Curr Opin Biotechnol. 2001 Apr;12(2):180-7. Review. PubMed PMID:11287235.
  22. Dorfman NA, Civin CI, Wunderlich JR. Susceptibility of adherent versus suspension target cells derived from adherent tissue culture lines to cell-mediated cytotoxicity in rapid 51Cr-release assays. J Immunol Methods. 1980;32(2):127-39. PubMed PMID: 7358991.
  23. Fogh J, Holmgren NB, Ludovici PP. A review of cell culture contaminations. In Vitro. 1971 Jul-Aug;7(1):26-41. Review. PubMed PMID: 5003037.
  24. Barry C. Buckland. Applications of cell culture for the improvement of human health. Cytotechnology. 2004;15:1-2.
  25. Robert N. Trigiano, dennis J. Gray. Plant Tissue Culture Concepts and Laboratory Exercises. Second edition. Nat Med 1997; 4; 402-5.
  26. T. Gaspar et al. Concepts in plant stress physiology. Application to plant tissue cultures Volume 37, Number 3 / July, 2002.
  27. Edwin F. George et al. Plant Propogation by Tissue Culture. (2008) 3rd edition, vol 1.
  28. Ashley Nixon. Plant Tissue Culture and Biotechnology. Volume 11, Number 1 / March, 1991.
  29. Harper MF, Sibley PE, Rowlands A, et al. Hormonal modulation of the growth of a new transplantable prostatic cellline in the nude mouse. Prostate 1985; 7: 369-76.
  30. Edelman, G. M., Murray, B. A., Mege, R-M., Cunningham, B. A., and Gallin, W, J. (1987.
  31. Chen, C., and Okayama, H. (1987). High efficiency transformation of mammalian cells by plasmid DNA. Mol.Cell. Biol. 7, 2745-2752.
  32. Harold N Trick et al. March 1997 Vol 3, No. 1. Plant Tissue Culture and Biotechnology.
  33. Klein KA, Reiter RE, Redula J, et al. Progression of metastic human prostate cancer to androgen independence in immunodeficient SCID mice. Nat Med 1997; 4; 402-8.
  34. The Department of Immunization, Vaccines and Biologicals. (2004). Manual for the virological investigation of polio. Retrieved April 17, 2010, from
  35. Cooper G. M. & Hausman R. E. (2000). The Cell: A Molecular Approach (2nd ed.). Bethesda, Md: Sinauer Associates.
  36. Thomas Gaspar et al. Volume 32, Number 4 / October, 1996. Plant hormones and plant growth regulators in plant tissue culture. “In Vitro Cellular & Developmental Biology - Plant”.
  37. . M. R. Ahuja. (1992). Micropropagation of Woody Plants.
  38. A. Mujib, J. Samaj (2006). Plant Cell Monographs. Somatic Embryogenesis.
  39. Elisa Eiseman, Sussane B. Haga. (1999). Handbook of Human Tissue Sources.
  40. Jan A. Mol, Roger A. Clegg. (1999). Biology of the Mammary Gland.
  41. Barbulovic-Nad I, Au SH, Wheeler AR. A microfluidic platform for complete mammalian cell culture. Lab Chip. 2010 Apr 15. [Epub ahead of print] PubMed PMID:20393662.
  42. Lee SH, Lillehoi HS, Hong YH, Jang SI, Lillehoi EP, Ionescu C, Mazuranok L & Bravo D.In Vitro Effects of Plant & Mushroom Extracts on Immunological Function of Chicken Lymphocytes & Macrophages PMID- 20461582.
  • John Wiley & Sons. (1998) :Applications of Plant Cell and Tissue Culture", Page 36-44, A Wiey-Interscience Publication., 1998.
  • S.S Bhojwani. (1990) "Plant Tissue Culture: Applications and Limitations", Page 86-90, 320, Elsevier Science Publishers B>V., 1990.

2010 Projects

Fluorescent-PCR | RNA Interference | Immunohistochemistry | Cell Culture | Electron Microsopy | Confocal Microscopy | Monoclonal Antibodies | Microarray | Fluorescent Proteins | Somatic Cell Nuclear Transfer