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  • 1. Cell Cultivations - involves cell elongation, cell wall - bound together by intercellular material to form tissue. ◦ Cytosol - the liquid around all of the floating structures between MICROBIAL CELL CULTIVATIONS invagination, distribution of nuclear ◦ Tissue: epithelium, connective tissue, muscle, & nerve. the nucleus and cell membrane. - Microorganisms are used in the production of pharmaceuticals, material, transverse cell wall - Epithelial tissue forms the covering or lining of all free body ◦ Chloroplast - site of photosynthesis in the plant cell, w/c contains industrial chemicals, enzymes, recombinant DNA technologies, formation, distribution of cellular surfaces, both external and internal. the green pigment chlorophyll (responsible for trapping the light for etc. material into two cells, and - Connective tissue, where cells are embedded in an extensive the production of carbohydrates) Microbial Cells separation into two new cells. intercellular matrix as either be solid, semisolid, & liquid. ◦ Nucleus - cell's control center and contains DNA for the protein ▪ All living organisms, classified as animal kingdom, plant ◦ Nutritional Req’ts: actively - Muscle cells are elongate & bound together into sheets or bundles translation and transcription. kingdom, (multicellular) and protist (Unicellular), (Haekel,1866). growing bacterial cells are 90% by connective tissue. ◦ Endoplasmic reticulum – is a network of tiny tubes that ▪ Protist, simple biological organism including algae, protozoa, water. - Muscles are responsible for most movement in higher animals interconnect the different parts of the cells. fungi, and bacteria. - phototrophs: organisms that capable of employing radiant energy. - Nerve cells, composed of a cell body (fibers, long thin extensions ◦ Ribosome - is concentrated on the surface of endoplasmic ▪ Unit structure: prokaryotes and eukaryotes. - chemotrophs: organisms that obtain energy for their activities and & nucleus), transmit impulses very rapidly. reticulum, which involved in the synthesis of proteins ▪ Prokaryotic cell: bacteria and blue-green algae, has two internal self-synthesis from chemical reactions that can occur in the dark. ▪ Suspension Cells: are nonanchorage-dependent cells from blood or ◦ Mitochondrion contains genetic material and enzymes important regions: cytoplasm and nuclear region (nucleoplasm) - autotrophs: organisms that use CO2 or CO3` as a carbon source. lymph fluids that do not require a surface to grow on. for cell metabolism. - is surrounded with a cell wall and a cell membrane. - heterotrophs: organisms which cannot use CO2 but glucose or - lymphocytes, derived from the lymphoid tissue & spherical. ▪ Plant Tissue Culture - refer only to those of unorganized aggregates ▪ Cell wall, protects the cell from external influences. amino acids as a sole carbon source. ▪ Anchorage-Dependent Cells: require a wettable surface for of cells - Types: Unorganized growth & organized growth ▪ Cell membrane (cytoplasmic membrane), a selective barrier ◦ Physical Conditions: temperature, gaseous environment, pH attachment and growth.(epithelial or fibroblast, widely used) - Unorganized growth, occurs frequently when pieces of whole between the cell interior & the external environment - the rate of microbial growth is temperature dependent. - microcarriers: microscopically small beads plants are cultured in vitro.(Callus cultures, amorphous cell - serves as the surface onto which other cell substances attached - Types of bacteria with respect to temperature: psychrophiles (-7 to Monoclonal Antibodies (identical antibody) aggregates; Suspension cultures, cell aggregates growing dispersed ▪ Cytoplasm has grainy dark spots (ribosomes: contains protein & 35°C: opt. temp, 20-30°C), mesophiles (7 to 45°C: opt. temp, - can be analyzed by the solid-phase assay techniques (ELISA, in liquid medium; Protoplast cultures, growth of protoplasts in solid ribonucleic acid (RNA) 30-40°C), or thermophiles (40 to 75°C: opt. temp, 45-60°C) enzyme linked immunosorbent assays; RIA, radioimmuno assays). or liquid medium). ▪ Ribosome is the site for protein - Types of bacteria with respect to O2: Aerobic bacteria (presence of - antibodies can bind to the antigens to eliminate foreign substances - Organized growth, occurs when organized plant parts are synthesis. free oxygen), anaerobic bacteria (absence of free oxygen), ◦ 5 classes of antibodies (immunoglobulin, Ig): G, A, M, D, and E transferred to culture medium with their structure preserved. (Root ▪ Nuclear region contains DNA (genetic Facultatively anaerobic bacteria (either absence or presence of free - Ig G, has a Y-shaped protein structure comprised of one pair of cultures, root tips taken from plants; Embryo cultures, embryos information that determines the oxygen), Microaerophilic bacteria (presence of minute quantities of heavy chain and light chain polypeptides, linked by disulfide bonds. removed from sterilized seeds, ovules, or fruits) production of proteins & other cellular free oxygen.) - antibody-secreting cells cannot be maintained in a culture medium. -- somatic embryos, embryos produced from cell culture technique. substances/structures). - optimum pH for bacterial growth: 6.5 - 7.5 Cell Fusion: Secondary Metabolite – classified into 3 major categories: alkaloids ▪ DNA fragments & low molecular ◦ Gram Reaction: Gram staining, differential staining technique that - myeloma cells, malignant tumor cells of the immune system. (crystalline, nitrogen-containing compds. which can be extracted by weight proteins – largest molecules to separates bacteria into gram-positive (purple) & gram-negative (red). - hybridoma, the fused cell of B-Iymphocytes with myeloma cells. use of acidic solutions), essential oils (mixtures of terpenoids), and cross the cell membrane. ▪ Fungi - plants devoid of chlorophyll & are unable to synthesize PLANT CELL CULTIVATIONS glycosides (phenolics, tannin, saponins, dye, etc.) ▪ Eukaryotic cell: plants, animals, protozoa, fungi, and algae. their own food. (yeast & molds) ◦ Secondary metabolites - serve as a chemical interface between the - Biotic elicitors are compounds or substances of microbial origin - has internal unit membrane systems that segregate the functional - Chlorophyll, a complex molecule which adsorbs visible light producing plants and their surrounding environment. - Abiotic elicitors are stress agents such as UV irradiation components of the cell. ◦ Yeasts: are unicellular organisms with spherical shape, 1-5 μm in - The technical rationale for using plant tissue is based on the unique CELL GROWTH MEASUREMENT - 1,000 to 10,000 times larger/complex than prokaryotic cells. width and 5-30 μm in length. (budding, its growth pattern) biochemical totipotency (ability to generate or regenerate a whole ◦ Balanced growth - growth during which a doubling of the biomass ◦ Nucleus - surrounded by a double membrane with pores 40 - 70μm - Saccharomyces cerevisiae, used in the manufacture of wine & beer organism from a part) of plant cells. is accompanied by a doubling of all other measurable properties of wide, containing chromosomes. and in the leavening of bread. ◦ Primary metabolites, produced in larger quantities than the the population such as protein, DNA, RNA, and intracellular water. - controls hereditary properties and all vital activities of the cell. ◦ Molds, a filamentous fungi (single cell ─ hyphae ─ mycelium) secondary - Types of chambers for counting cell number: hemocytometer, a ◦ Chromosomes - long, threadlike bodies found in the nuclei of cells, - Imp. Classes: Aspergillus and Penicillium metabolites that blood cell counting chamber with organisms of 3μm in diameter; which contain the genes arranged in nucleoproteins. ▪ Culture Media: cultivation, growth of microbial population in have specific Petroff-Hausser counting chamber, for use primarily with bacteria. ◦ Cytoplasm contains ribosome (large numbers of granules, artificial environment. metabolic Viable Plate Count: concentrated along the rough surface of the endoplasmic reticulum). ◦ Pure culture - culture that contains only one kind of functions. ◦ Viable cell - able to divide and form a colony. ◦ Endoplasmic reticulum - an irregular network of interconnected microorganism Plant Cells ◦ 2 ways of performing a plate count: spread plate method, a volume membrane-delimited channels. ◦ Mixed culture - contains more than one kind of microorganism. ◦ size : 20-40μm of no larger than 0.1 mL is spread over the agar surface; pour plate ◦ Mitochondria - contain the electron transport enzymes that utilize Steps for cultivating microorganisms: in diameter method, sample is mixed w/ melted agar & pour into a sterile plate. oxygen in the process of energy generation. (a) culture medium preparation, (b) sterilization, (c) inoculation &100-200μm Measurement of Cell Mass ◦ Vacuole & lysosome - organelles that serve to isolate various ◦ Types of culture media: long - absorbency is defined as the logarithm of the ratio of the intensity chemical reactions in a cell. - natural media (empirical/complex), used on the basis of - its structure of light striking the suspension to that transmitted by the suspension Microbial Nomenclature: binomial system experience similar to that of typical eukaryotic cells CELL IMMOBILIZATION - 1st word: genus - synthetic media (chemically defined), consist of dilute, reproducible - features: rigid wall, large vacuole, and presence of chloroplasts. - encouraging production of high yields of secondary metabolites - 2nd word: species name solutions of chemically pure, in/organic compounds. - surrounded by a cell wall (determines many features of the Bacillus: a small rod ◦ Sterilization: moist heat (steam under pressure) in an autoclave plant). Lactobacillus: a small milk rod - @15 psi & 121°C (15-20 mins for liquid media). ◦ middle lamella - outer layer of the cell wall (carbohydrate in Micrococcus: a small grainClostridium. a small spindle ◦ Inoculation - seeding of a culture vessel with the microbial material nature), contains heavy layer of pectin (serves as the glue to hold Pasteurella: after Louis Pasteur, Latinized (inoculum). one plant cell to an adjacent cell). Salmonella: after Daniel E. Salmon, Latinized ANIMAL CELL CULTIVATIONS ◦ cell membrane (cytoplasmic membrane) - inner layer of the wall, Saccharomyces: sugar fungus - mammalian cell culture technique employed to produce human thin & flexible & composed of protein and lipid. Bacteria - unicellular microscopic organisms growth hormones, interferon, plasminogen activator, viral vaccines, - regulates the movement of substances into and out of the cell - 1,500 known species occurring in all natural environments. and monoclonal antibodies, extracted from human cadavers. ◦ vacuole serves as a receptacle for waste metabolic products or - typical cell diameter: 0.5 to 1μm. - post-translational modifications: involve proteolytic cleavage, secondary plant substances. ◦ Shapes: cocci: spherical or ovoid subunit association, or a variety of additional reactions - occupies 90 percent of the volume of the cell for a mature plant bacilli: cylindrical or rod shaped (glycosylation, can protect proteins against proteolytic breakdown, cell spirilla: helically coiled maintain structural stability, & alter antigenicity). - is surrounded by a plasma membrane. - can protect cells & decrease problems related to shear forces. ◦ Growth Pattern: binary fission, asexual reproductive process. Animal Cells - are eukaryotic cells. - component: water, which contains dissolved solutes,