tissue culture hybridization

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tissue culture hybridization

  1. 1. jaisreenivasan
  2. 2. Contents <ul><li>Definition </li></ul><ul><li>History </li></ul><ul><li>Applications </li></ul><ul><li>Benefits </li></ul><ul><li>Procedure </li></ul><ul><li>Contamination </li></ul><ul><li>Disadvantages </li></ul><ul><li>Conclusion </li></ul>
  3. 3. Definition <ul><li>Tissue culture is a special type of asexual propagation where a very small piece of tissue (shoot apex, leaf section, or even an individual cell) is excised (cut-out) and placed in sterile (aseptic) culture in a test tube, petri dish or tissue culture container containing a special culture medium </li></ul><ul><li>Tissue culture is a way to maintain the viability of an animal or plant tissue outside the donor's body. </li></ul>
  4. 4. History <ul><li>In 1885  Wilhelm Roux  removed a section of the medullary plate of an embryonic chicken and maintained it in a warm saline solution for several days , establishing the basic principle of tissue culture </li></ul><ul><li>In 1907 Ross Harrison at the Rockefeller Institute cultured frog embryo nerve fibers and successfully demonstrated growth of the nerve fibers outside a living body or in culture mediums </li></ul><ul><li>Further investigations led to comprehensive protocols that allow for sterilized or contamination-free culture methods. </li></ul>
  5. 5. Applications -Medicine and Research <ul><ul><li>There are various applications of tissue culture in a wide array of scientific industries. </li></ul></ul><ul><ul><li>Cancer cells are analyzed in vitro to produce appropriate medicines that will eliminate the abnormal cells . </li></ul></ul><ul><ul><li>Skin grafting , wherein severely burned tissues are replaced with new ones using cultured cells . </li></ul></ul>
  6. 6. Applications -Plant Fiber Culture <ul><li>One of the most common types of tissue culturing is conducted in botanical and agricultural research . </li></ul><ul><li>Plant tissue is much easier to grow in a nutrient rich suspension than human or animal tissue, and with the right nutrients, different tissue samples can often be encouraged to develop as they would on a normal plant specimen. </li></ul>
  7. 7. Applications - Genetic Modification <ul><li>Plant tissues are also cultured when experimented with genetic modification. </li></ul><ul><li>Most gene modification on plants is conducted to alter a specific part of the plant and the way it grows. By isolating those tissues and growing them in a lab, scientists can quickly test their gene modification and spot any problems </li></ul>
  8. 8. Points to consider for Tissue Culture <ul><li>The culture medium contains a gel (agar) with the proper mixture of nutrients, sugars, vitamins and hormones, which causes the plant part to grow at very rapid rates to produce new plantlets. </li></ul><ul><li>A very specialized laboratory is required for tissue culture. All the procedures are done in a laboratory and special ventilated cabinet that is as sterile as an operating room. </li></ul>
  9. 9. Methods <ul><li>Biopsy </li></ul><ul><li>Swabs </li></ul><ul><li>These methods may involve suction, forceps or needle extractions. </li></ul><ul><li>Provide a lot of cells for testing or observation. </li></ul><ul><li>This is a painful and an invasive method of tissue collection. </li></ul><ul><li>The process includes using a wooden dowel or cotton swab and wiping it along the section of the body being tested </li></ul><ul><li>This removes the cells mechanically and deposits them onto the collection surface </li></ul><ul><li>This is a non-painful method of collection and limits invasive techniques. </li></ul>
  10. 10. Plant tissue culture <ul><li>These are the different stages of growth </li></ul>
  11. 11. Benefits <ul><li>Exact Duplication </li></ul><ul><li>Tissues harvested from a plant create an exact duplication, with the same genetic material and physical characteristics. </li></ul><ul><li>Timely Duplication </li></ul><ul><li>If a rare or slow-growing plant has not flowered or produced seed, tissue culture facilitates new plant development at any time of year. </li></ul>
  12. 12. Benefits <ul><li>Disease Avoidance </li></ul><ul><li>If a plant is diseased, tissue from small areas of the plant not inflicted by the disease can be taken to produce healthy, new plants. Moreover, the isolated sterile environment of the tissue culture itself can prevent spread of diseases and insect pests. </li></ul><ul><li>Economics </li></ul><ul><li>Indoor laboratories can produce huge and repeated numbers of plants year round in small spaces, a huge advantage over traditional propagation techniques like sowing and rooting cuttings </li></ul>
  13. 13. Tissue culture <ul><li>A disease free plant </li></ul>
  14. 14. Procedure -Cell Types and Lines <ul><li>Primary cells come directly from a living organism and they can originate from any organ. These cells are difficult to culture because they have limited ability to divide, making their life spans short. </li></ul><ul><li>Finite cell lines also come directly from animal tissues and are difficult to propagate. These cells can be cultured over several generations </li></ul><ul><li>Continuous cell lines come from an action called transformation whereby their genetics have spontaneously changed in culture. These cells are easy to propagate because they have an almost indefinite lifespan </li></ul>
  15. 15. Procedure - Culturing cells <ul><li>The method of cell culture will be different for each cell type </li></ul><ul><li>Most cells are kept frozen </li></ul><ul><li>Frozen cells must be thawed in a 37 o C warm water bath quickly in order to prevent unnecessary cell death. In addition, the growth media that is used in the plates should also be warmed prior to use. After thawing, transfer the cells to a culture dish with the prescribed amount of nutrient media and incubate for 24 hours. This time period allows the cells to stick to the bottom of the dish so that the media can be removed the next day and replaced with fresh media. </li></ul>
  16. 16. Procedure - Observation and Maintenance <ul><li>Monitor your cells daily by looking at them under the microscope. Look for signs of failure, such as many floaters. </li></ul><ul><li>Use the suction tip in the tissue culture hood to evacuate the media and dead cells and then refresh with new media. </li></ul><ul><li>Healthy cells should replicate. When the bottom of the plate is almost fully covered with cells, it is time to split them. </li></ul><ul><li>This is done by washing the cells with a buffer solution and then detaching the cells from the bottom of the plate using media and a pipette or scraping </li></ul>
  17. 17. Contamination - Chemical Contamination <ul><li>Chemical contamination is the presence of any non-living substance in the cell culture that causes adverse effects to the cells </li></ul><ul><li>This include impure media, serum or even water which may contain undesirable endotoxins </li></ul><ul><li>Exposure to too much fluorescent lighting can alter the chemical composition of the media negatively. </li></ul><ul><li>Chemical contaminants could also come from unclean storage vessels. </li></ul>
  18. 18. Contamination - Cross Culture Contamination <ul><li>Cross culture contamination occurs when a different cell type is inadvertently introduced into the cells that are being cultured. </li></ul><ul><li>Cross culture contamination voids the experiment because the effect of the contaminating cells is unknown. </li></ul>
  19. 19. Contamination - Bacteria, Fungi or Yeast <ul><li>These microorganisms are the most common cell culture contaminators </li></ul><ul><li>Visual indicators of contamination include media color change </li></ul><ul><li>Presence of non-cellular material </li></ul><ul><li>Cell vacuolization </li></ul><ul><li>Mycoplasma (parasite)are very small bacteria-like organisms that are troublesome contaminators </li></ul><ul><li>It can’t be detected by these indicators </li></ul>
  20. 20. Disadvantages <ul><li>Tissue culture has both positive and negative aspects. </li></ul><ul><li>Cost Factor </li></ul><ul><ul><li>Plant tissue culture experiments involve high cost because of the expensive machinery required. </li></ul></ul><ul><li>Complexity </li></ul><ul><ul><li>Plant tissue culture is a complex scientific analysis. The procedure is multifaceted depending on the type of plant being incubated. </li></ul></ul><ul><ul><li>the plants all have the same genetic material, genetic diversity is reduced </li></ul></ul>
  21. 21. Conclusion <ul><li>Tissue culturing is an important advancement in biological research that allowed investigators to analyze the growth, development and function of cells. </li></ul><ul><li>To carry out tissue culturing effectively, several factors such as light, temperature, type of culture medium and pH must be strictly maintained </li></ul><ul><li>It has both advantages and disadvantages. So it is in our hands to use it carefully </li></ul>

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