1、Cell And Tissue Culture,Mammalian Cells Advanced Higher Biology,Why is it useful?,Gene manipulationCulturing mammalian cells for cancer studiesProducing new plants through tissue culture,What do you need to do it?,Source of cell material-freshly prepared-stock of cell line-bacterial culture,Suitable
2、 container,Simple flaskSophisticated fermenter with computer-controlled monitoring,Growth medium,Glucose Water Amino acids Salts,Opportunity for Gas Exchange,OxygenCarbon dioxide,Animal serum,Foetal Bovine SerumEssential for animal cell proliferation5% - 10% of growth media,Indicator,Waste products
3、causes change in pHUse indicator like phenol red Changes from red to yellow,Control of Temperature and pH,37.5 OCpH 7.5,Method for Measuring Cell Growth,Counting cell numbers in culture (haemocytometer)Measure optical density in spectrophotometer,Sterilisation,AntibioticsSterilisation,Cells are eith
4、er.,Anchorage dependantAnchorage - independant,Anchorage independant cells,Cells associated with body fluid-blood cellsGrown in suspensionWill eventually need subculturing,Anchorage dependant cells,Most animal derived cellsAdhere to bottom of a flask and form a monolayerEventually cover entire surfa
5、ce of substratum (confluence)Proliferation then stopsNeed to subculture cells at this point (remove to fresh medium)Proliferation can begin again,2 main categories of animal cell cultures.,Primary cultureContinuous cell line,Primary Cultures,Taken from fresh tissueLimited life span in cultureTreated
6、 by proteolytic enzyme (Trypsin)Separate into single cells-epithelial cells-fibroblasts,Continuous Cell Line,Derived from humans Been transformed-lose sensitivity to factors associated with growth controlProduce immortalised cell linesCell lines are neoplasticOften lose their anchorage-dependence-as
7、sociated with an altered xsome patternMore easily cultured,Why do it?,Particular cells can be isolated and cloned-isolation of mutant cells-investigate cell growth-produce hybrid cells that have applications in biotechnologyProduce important pharmaceuticals-vaccines-hormones,This powerpoint was kindly donated to http:/ is home to over a thousand powerpoints submitted by teachers. This is a completely free site and requires no registration. Please visit and I hope it will help in your teaching.,
