Catálogo de Células

0100 - HeLa

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BCRJ Code 0100
Cell Line HeLa
Species Homo sapiens
Vulgar Name Human
Tissue Cervix
Cell Type Epithelial
Morphology Epithelial
Disease Adenocarcinoma
Growth Properties Adherent
Sex Female
Age/Ethinicity 31 Year / Black
Applications These cells are a suitable transfection host. This cell line can be used to screen for Escherichia coli strains with invasive potential.
DNA 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
Virus Succeptility: Human adenovirus 3 Encephalomyocarditis virus Human poliovirus 1 Human poliovirus 2 Human poliovirus 3
Products Keratin
Biosafety 2
Addtional Info The cells are positive for keratin by immunoperoxidase staining. HeLa cells have been reported to contain human papilloma virus 18 (HPV-18) sequences. P53 expression was reported to be low, and normal levels of pRB (retinoblastoma suppressor) were found.
Culture Medium Dulbecco's Modified Eagle's Medium (DMEM) with 1% non-essential amino acids, 2 mM L-glutamine, 1 mM sodium pyruvate, 1.0 g/L glucose and 10% of bovine serum.
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. T-75 flasks are recommended for subculturing this product. Remove and discard culture medium. Briefly rinse the cell layer with PBS without calcium and magnesium to remove all traces of serum that contains trypsin inhibitor. 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°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010.
Subculturing Medium Renewal 2 to 3 times per week
Subculturing Subcultivation Ratio 1:2 to 1:6
Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C
Cryopreservation 95% FBS + 5% DMSO (Dimethyl sulfoxide)
Thawing Frozen Cells SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris. 1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes). 2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions. 3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes. 4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). 5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line). NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
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Bacteriol. 178: 6281-6287, 1996. PubMed: 8892830 Mansky LM. The mutation rate of human immunodeficiency virus type 1 is influenced by the vpr gene. Virology 222: 391-400, 1996. PubMed: 8806523 Dobbelstein M, Shenk T. Protection against apoptosis by the vaccinia virus SPI-2 (B13R) gene product. J. Virol. 70: 6479-6485, 1996. PubMed: 8709286 Churchill MJ, et al. The rev-responsive element negatively regulates human immunodeficiency virus type 1 env mRNA expression in primate cells. J. Virol. 70: 5786-5790, 1996. PubMed: 8709194 Goodrum FD, et al. Adenovirus early region 4 34-kilodalton protein directs the nuclear localization of the early region 1B 55-kilodalton protein in primate cells. J. Virol. 70: 6323-6335, 1996. PubMed: 8709260 Kolanus W, et al. alphaLbeta2 integrin/LFA-1 binding to ICAM-1 induced by cytohesin-1 a cytoplasmic regulatory molecule. Cell 86: 233-242, 1996. PubMed: 8706128 Gan W, Rhoads RE. 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Human immunodeficiency virus type 1 cell cycle control: Vpr is cytostatic and mediates G2 accumulation by a mechanism which differs from DNA damage checkpoint control. J. Virol. 70: 2324-2331, 1996. PubMed: 8642659 Roller RJ, et al. Structure and function in the herpes simplex virus 1 RNA-binding protein US11: mapping of the domain required for ribosomal and nucleolar association and RNA binding in vitro. J. Virol. 70: 2842-2851, 1996. PubMed: 8627758 Jang SI, et al. Activator protein 1 activity is involved in the regulation of the cell type-specific expression from the proximal promoter of the human profilaggrin gene. J. Biol. Chem. 271: 24105-24114, 1996. PubMed: 8798649 Anderson SM, et al. Intercellular transfer of a glycoslphosphatidylinositol (GPI)-linked protein: release and uptake of CD4-GPI from recombinant adeno-associated virus-transducted HeLa cells. Proc. Natl. Acad. Sci. USA 93: 5894-5898, 1996. PubMed: 8650189 Dittrich E, et al. 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Comprehensive and definitive molecular cytogenetic characterization of HeLa cells by spectral karyotyping. Cancer Res. 59: 141-150, 1999. PubMed: 9892199 Dentistry--Preclinical evaluation of biocompatibility of medical devices used in dentistry--Test methods for dental materials. Geneva (Switzerland):International Organization for Standardization/ANSI;ISO ISO 7405:1997.
Depositors Departamento de Virologia, Instituto de Microbiologia, UFRJ, Rio de Janeiro.
ATCC CCL-2
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