hct-116、hela、MEFs细胞参数介绍.doc

HELAHeLa (ATCC® CCL-2™)ATCC：Hela 细胞，源于黑人 31 岁女性，子宫颈腺癌，是一种附着型上皮细胞，要求存于液氮中。These cells are a suitable transfection host.This cell line can be used to screen for Escherichia coli strains with invasive potential.Biosafety Level：（2 [Cells contain human papilloma virus]Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.）Complete Growth Medium：The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.Subculturing（接种）：Volumes used in this protocol are for a 75 cm2flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. CorningT-75 flasks (catalog #430641) are recommended for subculturing this product.1.Remove and discard culture medium.2.Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.3.Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37C to facilitate dispersal.4.Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.5.Add appropriate aliquots of the cell suspension to new culture vessels.6.Incubate c...