CCD 1106 KERTr

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货期:

编号:TS212468

品牌:Testobio

产品名称：	CCD 1106 KERTr
商品货号：	TS212468
Organism：	Homo sapiens, human
Tissue：	skin
Cell Type：	keratinocyte; human papillomavirus 16 (HPV-16) E6/E7 transformed
Product Format：	frozen
Morphology：	epithelial
Biosafety Level：	2 Cells may contain the Human Papilloma viral (HPV) sequences 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.
Age：	133 days gestation
Storage Conditions：	liquid nitrogen vapor phase
Derivation：	Passage 2 cells were transformed with a retrovirus vector (LXSN16E6E7 produced by ATCC CRL-2203) in the presence of polybrene.
Antigen Expression：	epithelial specific antigen
Oncogene：	E6/E7 +
Genes Expressed：	E6/E7 +, epithelial specific antigen
Comments：	This line stains positively with antibody for cytokeratin. After 50 population doublings, the cells continue dividing and retain cuboidal morphology. E6E7 sequences were detected by PCR in cells at passage 18. Major Histocompatibility Complex class I or II molecules were not expressed on these cells, but PCR analyses revealed presence of the genes for directing synthesis of HLA antigens. Epithelial specific antigen was detected using antibody produced by the Ep16 hybridoma (ATCC HB-155).
Complete Growth Medium：	These cells are grown in Keratinocyte-Serum Free Medium (Gibco 17005-042) with added Keratinocytes Supplements (Gibco 37000-015) including Bovine Pituitary Extract (BPE; Gibco 13028-014) and human recombinant epidermal growth factor (EGF; Gibco 10450-013) further supplemented with:xa0 Additional 35 ng/mL human recombinant epidermal growth factor (EGF; BD cat# 354052).xa0 Do not filter the complete medium. xa0 This medium is formulated for use with a 5% CO₂ in air atmosphere.
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. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mMxa0EDTA solution 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. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium.xa0 Add appropriate aliquots of cell suspension to new culture vessels. Place culture vessels in incubators at 37°C. Subculture Ratio: 1:3 to 1:5 Medium Renewal: Twice a week. Note:xa0Add fresh medium twice per week Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.
Cryopreservation：	Freeze Medium: Hams F12 medium 85%; DMSO, 5%; FBS 10% Storage Temperature: Liquid nitrogen vapor phase
Culture Conditions：	Temperature: 37°C
Name of Depositor：	L Vilner
Passage History：	E6E7 sequences were detected by PCR in cells at passage 18. Passage 2 cells were transformed with a retrovirus vector ( LXSN16E6E7 produced by ATCC CRL-2203) in the presence of polybrene.
References：	Zabrenetzky V, et al. The isolation, immortalization and characterization of human fetal keratinocytes. In Vitro Cell. Dev. Biol. 33: Part II, p. 34A, 1997.

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CCD 1106 KERTr

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产品名称：	CCD 1106 KERTr
商品货号：	TS212468
Organism：	Homo sapiens, human
Tissue：	skin
Cell Type：	keratinocyte; human papillomavirus 16 (HPV-16) E6/E7 transformed
Product Format：	frozen
Morphology：	epithelial
Biosafety Level：	2 Cells may contain the Human Papilloma viral (HPV) sequences 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.
Age：	133 days gestation
Storage Conditions：	liquid nitrogen vapor phase
Derivation：	Passage 2 cells were transformed with a retrovirus vector (LXSN16E6E7 produced by ATCC CRL-2203) in the presence of polybrene.
Antigen Expression：	epithelial specific antigen
Oncogene：	E6/E7 +
Genes Expressed：	E6/E7 +, epithelial specific antigen
Comments：	This line stains positively with antibody for cytokeratin. After 50 population doublings, the cells continue dividing and retain cuboidal morphology. E6E7 sequences were detected by PCR in cells at passage 18. Major Histocompatibility Complex class I or II molecules were not expressed on these cells, but PCR analyses revealed presence of the genes for directing synthesis of HLA antigens. Epithelial specific antigen was detected using antibody produced by the Ep16 hybridoma (ATCC HB-155).
Complete Growth Medium：	These cells are grown in Keratinocyte-Serum Free Medium (Gibco 17005-042) with added Keratinocytes Supplements (Gibco 37000-015) including Bovine Pituitary Extract (BPE; Gibco 13028-014) and human recombinant epidermal growth factor (EGF; Gibco 10450-013) further supplemented with:xa0 Additional 35 ng/mL human recombinant epidermal growth factor (EGF; BD cat# 354052).xa0 Do not filter the complete medium. xa0 This medium is formulated for use with a 5% CO₂ in air atmosphere.
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. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mMxa0EDTA solution 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. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium.xa0 Add appropriate aliquots of cell suspension to new culture vessels. Place culture vessels in incubators at 37°C. Subculture Ratio: 1:3 to 1:5 Medium Renewal: Twice a week. Note:xa0Add fresh medium twice per week Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.
Cryopreservation：	Freeze Medium: Hams F12 medium 85%; DMSO, 5%; FBS 10% Storage Temperature: Liquid nitrogen vapor phase
Culture Conditions：	Temperature: 37°C
Name of Depositor：	L Vilner
Passage History：	E6E7 sequences were detected by PCR in cells at passage 18. Passage 2 cells were transformed with a retrovirus vector ( LXSN16E6E7 produced by ATCC CRL-2203) in the presence of polybrene.
References：	Zabrenetzky V, et al. The isolation, immortalization and characterization of human fetal keratinocytes. In Vitro Cell. Dev. Biol. 33: Part II, p. 34A, 1997.