Cell Cultures - PDF

CELL CULTURES: It is the growth and proliferation of cellular models under controlled conditions. HISTORY OF CELL CULTURE 1907- Frog embryo nerve fiber outgrowth by Harrison. 1943- Development of mouse lymphocyte cell line by Earle, et al. • Cell culture is performed on 1952- Development of HeLa ce...

CELL CULTURES: It is the growth and proliferation of cellular models under controlled conditions. HISTORY OF CELL CULTURE 1907- Frog embryo nerve fiber outgrowth by Harrison. 1943- Development of mouse lymphocyte cell line by Earle, et al. • Cell culture is performed on 1952- Development of HeLa cell line by Gey, et al. 1955- Development of defined cell culture media by Eagle. artificial media prepared by mixing purified components or complex 1961- Demonstration of the finite lifespan of normal human organic solutions, cells by Hayflick and Moorhead. 1964- Discovery of pluripotency of embryonic stem cells by Kleinsmith and Pierce. 1976- Totipotency of embryonic stem cells described by Illmensee and Mintz. 2007- Use of viral vectors to reprogram adult cells to embryonic state (induced pluripotent stem cells) by Yu et al. 2008 and beyond- Era of induced pluripotent stem cells – promises and challenges. • Using equipment to maintain the appropriate physico-chemical conditions and on containers that isolate them from A culture medium consists of four elements 1. The nature of the substrate or growing phase. 1 .- Adherent: The majority of cells maintained in culture derived from tissue disintegration or tumors formed by adherent cells and maintain this characteristic: they need to adhere to the substrate to be maintained. (derived from organs: muscle, liver, nerve cells ...) 2 .- Suspension: The suspension cultures often coincide with those of cells that "in vivo" are circulating, overall blood cells, immune cells. Lab equipment will depend on this characteristic 2. The physico-chemical and physiological conditions of the medium 3. The nature and composition of the gas phase 4. Incubation conditions, specially humidity and temperature Cell Culture • Gaseous phase. The most significant components of the gas phase are oxygen and carbon dioxide. • Physical properties. Environmental characteristics are: pH, osmolarity, temperature, viscosity and surface tension. • Physiological conditions. They refer to the composition of the medium. The main difficulty in the establishment of cell lines is to obtain adequate nutritious media to be able to replace the "natural" environment such as embryonic extracts, protein hydrolysates or sera. Some of the main media used are: Basal Medium Eagle (BME), Eagle's Minimum Essential Medium (MEM), RPMI 1640 and MEM modified by Dulbeco Medium (DMEM). BIOCHEMICAL COMPOSITION OF CULTURE MEDIUM EXAMPLE: DMEM - Bicarbonate buffer, -à Maintenance of pH Salts, pyruvate, glucose, aas, vitamins…-à Nutrients (Primary cultures have more requirements) - Fetal Bovine Serum (5-15%)à Growth factors-à Proliferation - Antibiotics/Antimicotics (10%) à sterility - Phenol red, to observe medium acidification--à Control of the general status of the culture