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Lecture 1 Handout -Cell Cultures.docx

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Biology 2382B

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Cell Cultures Pg 3, 395-398, 906, 911, 915-916, 1107-1113 Carbon stores in bacteria is almost equal to the amount Single cell cultures proliferate into colonies, containing thousands of genetically identical cells. in plants. Euks contain a nucleus and internal membranes tht Primary Cell Cultures and Cell Differentiation enclose organelles. Skin, kidney, liver tissues and embryos are used to establish primary cell cultures. The cytoplasm is the region between the P.Membrane and the nucleus. It holds the cytosol and organelles. Prep for the cultures mean the cell-cell & cell-matrix interactions must be broken: Euks comprise both plant and animal kingdoms; including uni/multicellular fungi and protozoans. -Treat w/ protease & a cation chelator… Euk cells are 10-100 µm across in width.  The chelator depletes the medium of usable Ca & Mg ions that inactivates CAMs Culture of Animal Cells Req Mediums and Special Solid  CAMs that aren’t calcium dependent need to Surfaces be proteolyzed (trypsin, collegnase or both) Fibroblasts are the predominant cells in connective Isolated animal cells are placed in culture mediums. Kept in incubators where temperature, humidity and tissue & produce ECM components like collagen that atmosphere is controlled, they have added antibiotics anchor cells to a surface. to reduce bacterial/fungal contamination. Fibroblasts in culture divide more rapidly than other Requirements of Media: cells in a tissue & are dominant cell type in a primary culture unless precautions are taken. 1. 9 essential a.a. (cant be synthesized) Vertebrate embryos contain myoblasts, muscle cell 2. 3 other a.a. synthesized only by specialized cells 3. Vitamins, salts, glucose precursors that divide & migrate to other sites in the 4. Serum ( fluid remaining after blood plasma embryo. At some point: removed) 1. Serum has protein factors that are needed for the 1. Align w/ each other proliferation (Rapid reproduction of a cell) of 2. Cease division 3. Fuse into multinucleate cell: Myotubes mammalian cells in culture. Myotubes induce synthesis of dozens of muscle specific The protein factors include insulin, transferrin (supplies iron bioaccesibly), and multiple growth factors. proteins req’d for further muscle development & function. There are special cases where cells require protein growth factors not found in serum (p 396): Mononucleated Satellite cells are in adult muscle cells & 1. Red Blood Cell Progenitors fuse to form myotubes also. Simutaenously, they induce 2. T Lymphocytes synthesis of muscle specific proteins. Bacterial and yeast cells can be grown in suspension, In the proper medium, satellite cells fuse & differentiate but animal cells only grow on a solid surface… into muscle cells. CAMs (cell adhesion molecules) are important because they bind cells to other cells or proteins, like collagen. Fig 9.30 pg 397 Cells secrete ECM components that adhere to glass or plastic dishes/flasks. Cell Cultures Pg 3, 395-398, 906, 911, 915-916, 1107-1113 Cell Cultures & Strains have a Finite Lifespan After cells are cultures, the adherent ones divide a finite number of times & then cease growing – Cell Senescence. Around 50 times before they cease growth. Regardless of a limited lifetime, if maintained, a single culture can be studied through many generations. A lineage of cells originating from an initial primary culture is a Cell Strain. Prolonged growth by cells are derived from tumors. Rare cells in a population of primary cell cultures undergo transformations & therefore can grow indefinitely. Cell cultures w/ an indefinite lifespan are considered Normal cells can be differentiated into stem cells by immortal & are called Ce
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