Posted at 10.09.2018
The reason for the experiment is to compare and contrast normal cell and transformed cell. Students compare normal and transformed cell after taking a look at it under the microscope. The purpose of the test is to find the characteristics between normal and altered cell. Each college student will observe the morphological differences, as well as the differing progress patterns of these cell types when cultivated in culture. Normal skin cells have a condition of normal style and you will find some irregularity (1). If you perform to get ready a normal cell, whenever a cell grows and when cell touches then your growth ceases by contact inhibition. Transform cells (Cancers or Tumor cells) have a condition that is irregular and in the nucleus they have multiple nuclei that lead it to be abnormal (1). In abnormal cells there is no contact inhibition and the cells grow multiple level which contributes to tumor or cancer cell (2). In abnormal skin cells, Metastasis has malignancy cells copy to other part of the body therefore you see long membrane extension. Abnormal cell has "large cells" that includes a cell that has multiple nuclei and it looks swollen (1). You can find few things that cause normal skin cells to go to tumor cells. The first thing is Environmental Carcinogens which release chemicals in the air. The second is UV radiation. The third is Retroviruses which includes Hepatitis C that produces liver tumors. The fourth is HPV virsus which causes Cervical tumors.
The staining techniques were the same for normal and transformed cell. The group received one coverslip with each cell type fastened. The coverslips comprised markings which when focused properly by the group this implies the slide of the coverslip to which the cell is attached. The group dealt with the coverslip very carefully because the coverslips were fragile. The group then grasped the coverslip with a forcep and dipped it into stain one for only 1 second. The group dipped the coverslip into stain one once more. The group then transferred the coverslip immediately into stain two for only one second and repeated this step once more. The group prevented overstaining of the coverslips. The group then rinsed the coverslip with distilled normal water for few seconds to clean off any residual stain. The coverslip was then air dried out completely. The group blowed the coverslip softly to speed up the procedure for permount to be added to the microscope glide. Following the coverslip was completely dried the group took a wooden adhere to add one to two drops of permount to the middle of the microscope glide. The group then put the coverslip cell area down onto the permount. Following the coverslip was added the group applied lightly pressure to the coverslip to equally spread the permount between your slip and coverslip. The group averted air bubbles and later allowed the permount to dried up on the slide. The group detected the cells under a microscope with low and high power. In the reduced electricity the group could see the condition, growth habits, and cell circulation. In high vitality the group could view the nuclear, nucleolar figures, chromosomes, and cytological aberrations.
In L929 of the transform cell they have various cell figures characteristics. Due to the migration over the coverslip during expansion, the cell membrane extends in various guidelines. The membrane "ruffling" is characteristics of migratory cells and skin cells internalizing droplets of medium by pinocytosis. Note the large cell in the center of the photo on low power. This abnormality may result from the actual fact that the skin cells are cultivated on manufactured environment (plastic). The existence of the gigantic cell is an sign of problems in the cell culture environment unless the rate of recurrence of the cells increases above normal level. The multiple nuclei result from the loss of coordination between karyokinesis and cytokinesis, thus resulting in an elevated quantity of nuclei per cell. The cell in the heart of the photograph of high power which includes long membrane extension shows that it probably was considering migration when it was fixed. You can also note the variety of the cell patterns in the photo. You can even take note the binucleated cell on the lower right corner of low ability. This isn't rare for that one cell type.
In IMR 90 the expansion patterns of the skin cells were mentioned. IMR 90 is much bigger than the L929 cell so you can see the orientated expansion pattern. As shown in high power the nucleus was noticed in each cell. The nucleus is elliptical. The difference is size probably represents the difference in cell cycle positions. Whenever a confluent monolayer of skin cells is made in the culture flask they'll stop growing and migrating. Alternatively, most but not all transformed skin cells are not contact inhibition and can continue to accumulate on one another when confluency is reached. L929 usually will not continue to expand once confluency is reached. Contact inhibition is the cessation of mobile section when the skin cells have covered the whole surface of the growing vessel. That is a property common to all normal cells plus some transformed cell. An example of an agent (natural or environmental) which can bring about the forming of a changed cell is vitro (clear plastic). The abnormality may result from the fact that the cells are grown with an manufactured environment (clear plastic) which is also known as massive cell. Two distinctive morphological differences between your IMR-90 cells and the L-929 skin cells, one is progress style; the diploid human being lung fibroblast cell, IMR 90, is a lot larger than the L929 cell. The second reason is the difference in proportions probably represents dissimilarities in cell routine positions. "Giant" cells are a anomaly within some cell lines when development in culture and that triggers it to create. Giant skin cells form because when the cell expansion persists in the absence of cell division. Huge cells are reproductively deceased and metabolically alive. The purpose and action of trypsin is proteolytic enzyme which digests various extracellular proteins which bind cells to one another as well as binding skin cells to the tissues culture flask. Over trypsinization can lead to the destruction of skin cells as it begins to absorb the plasma membrane protein. The action of trypsin can be inhibited with the addition of serum following trypsinization. The serum contains an anti-trypsin agent which inactivates the enzyme. Aneuploidy is the problem which are present when the nucleus of your cell will not contain an exact multiple of the haploid number of chromosomes; a number of chromosomes being symbolized more or less times than the others. The chromosomes may or might not show rearrangements. I'd expect IMR 90 to get this condition because IMR 90 does not have exact multiple of haploid amount of chromosomes and other characteristics. In vivo imply existing or completed in the living organism, e. g. in a test or test. In vitro means in an artificial environment alternatively than inside a living organism, e. g. in a test pipe. When one examines a specimen with a microscope low ability should be utilized first because you can see the skin cells better and you could identify few things. In high ability you are zoomed in to the cell and also you go into more depth of the picture. It's important to do cell culture work using aseptic approach because cell culture is employed to denote the growing of skin cells in vitro like the culture of solitary cells and aseptic approach performs lab manipulation in the absence of fungi, bacteria, infections, or other microorganisms. When a researcher isolated a inhabitants of cells many years back and has prolonged to keep that cell range through subculturing, I would expect this cell range to be normal because cell range is a cell culture from a single cell and subculturing is a secondary biological culture. The contaminants rate is low and you keep taking the culture from a 100 % pure one so you would have normal skin cells.
Tumor cells vary from normal cells in several basic ways. The foremost is the separation of normal cells that is securely regulated by special cell alerts. With tumor skin cells the signals are no more produced or perhaps they are no longer received. Research skin cells tend to be able of getting rid of the cells from a person and growing them in a sterile dish with the nutrition required for their success (2). Growing cells is termed as "cell culture". The normal cells grow until the bottom with their dish is protected with the cell in culture. If the density is reached, they stop dividing because there is forget about space (2). If one cell dies another one divides to complete the space. Normal cells will split a certain quantity of times after the division process prevents (2). There are a certain programmed variety of generations which may be produced and then there is absolutely no more dividing (1). Finally, the whole culture will pass away.
Tumor skin cells will divide again and again and in culture it'll become a piled-up mess of cells. It is as though tumor cells lose the capability to follow the rules and they separate (proliferate) uncontrollable (2). A second major difference between normal skin cells and tumor skin cells is that normal cells perform a particular function for your body. Normal skin cells have specialized manners and serve as an objective. Normal cells extracted from different tissue have different appearances. Tumor skin cells have another type of appearance than normal cells taken from the muscle they are derived from. This is due to the fact they have lost their specialized function (2).
Gershman, H. , Drumm, J. , and Culp, L. Sorting out of normal and virus-transformed cells in mobile aggregates. The Journal of Cell Biology. 1976; 68: 276-286.
Woynarowska, B. , and Woynarowski, J. Preferential targeting of apoptosis in tumor versus normal cells. Biochimica et Biophysica Acta. 2002; 1587: 309-317.