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HEp-2 (ATCC® CCL-23™)

Organism: Homo sapiens, human /

Tissue: HeLa contaminant /

Disease: Carcinoma Tissue HeLa contaminant

Product Format frozen

Morphology epithelial Culture Properties adherent

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.

Disease Carcinoma

Storage Conditions liquid nitrogen vapor phase

Karyotype Occasional polyploids. Several marker chromosomes were observed along with frequent minutes, and often 2 large chromosomes with subterminal centromeres.HeLa Marker Chromosomes: One copy of M2, two-four copies of M3 and one copy of M4 as revealed by G-banding patterns.

Derivation Cells of this line contain HeLa marker chromosomes, and were derived via HeLa contamination. This line was originally thought to be derived from an epidermoid carcinoma of the larynx, but was subsequently found, based on isoenzyme analysis, HeLa marker chromosomes, and DNA fingerprinting, to have been established via HeLa cell contamination. The cells are positive for keratin by immunoperoxidase staining.

HeLa Markers Y

Genes Expressed keratin,The cells are positive for keratin by immunoperoxidase staining.

Cellular Products keratin Virus Susceptibility Human adenovirus 3 Human poliovirus 1

Vesicular stomatitis virus

Comments Cells of this line contain HeLa marker chromosomes, and were derived via HeLa contamination. This line was originally thought to be derived from an epidermoid carcinoma of the larynx, but was subsequently found, based on isoenzyme analysis, HeLa marker chromosomes, and DNA fingerprinting, to have been established via HeLa cell contamination. The cells are positive for keratin by immunoperoxidase staining. ATCC confirmed this cell line is positive for the presence of human papilloma viral DNA sequences via PCR.

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 Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

1，Remove and discard culture medium,

2，Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM 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 37℃ 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 cultures at 37℃.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:10 is recommended

Medium Renewal: 2 to 3 times per week

Cryopreservation

Freeze medium: culture medium 95%; DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

Culture Conditions

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37℃

STR Profile Amelogenin: X CSF1PO: 9,10 D13S317: 12,13.3 D16S539: 9,10 D5S818: 11,12 D7S820: 8,12 THO1: 7 TPOX: 8,12 vWA: 16,18

Isoenzymes G6PD, A