VK2/E6E7細(xì)胞

年限： 32 years *****

細(xì)胞形態(tài)： 上皮樣

生長狀態(tài)： 貼壁生長

組織來源： mucosa

運輸方式： 凍存運輸

數(shù)量： 大量

細(xì)胞類型： 其他細(xì)胞類型

是否是腫瘤細(xì)胞： 0

物種來源： 人

器官來源： 其他

ATCC Number： CRL-2616?

VK2/E6E7細(xì)胞Designations: VK2/E6E7

Depositors: D Anderson, RN Fichorova, JG Rheinwald

Biosafety Level: 2 [Cells contain human Papilloma viral sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens deposited as human

Morphology: epithelial

Source: Organ: vagina

Tissue: mucosa

Cell Type: epithelialHPV-16 E6/E7 transformed

Cellular Products: cytokeratins 8 (CK8), 10 (CK10), 13 (CK13), 18 (CK18) and 19 (CK19) [52983]

macrophage colony-stimulating factor (M-CSF); transforming growth factor beta1; interleukin 8 (IL-8); prostaglandin E2; the secretory leukoproteinase inhibitor; polymeric immunoglobulin receptor [52984]

Permits/Forms: VK2/E6E7細(xì)胞In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

Restrictions: The line is available with the following restrictions: 1. This cell line was deposited at the ATCC by Dr. Deborah Anderson of The Brigham and Women's Hospital, Inc. and is provided for academic research purposes only. Neither the cell line nor products derived from it may be sold or used for commercial purposes, including screening of compounds. Nor can the cells be distributed to third parties for purposes of sale, or producing for sale, cells or their products. The cells are provided as a service to the research community. They are provided without warranty of merchantability or fitness for a particular purpose or any other warranty, express or implied. 2. All products derived from the cells or genetically altered forms of the cells cannot be commercialized without express written permission from the The Brigham and Women's Hospital, Inc. 3. Any proposed commercial manufacture, use, or sale of the these cells, or their products must first be approved and the terms for such commercial manufacture, use, or sale must first be negotiated with TheBrigham and Women's Hospital, Inc., Office of Corporate Sponsored Research and Licensing, 75 Francis Street, Boston, MA 02115 (617) 525-6010.

Isolation: Isolation date: 1996

DNA Profile (STR): Amelogenin: X

CSF1PO: 10, 11

D5S818: 9, 10

D13S317: 9, 12

D7S820: 10, 11

D16S539: 9

vWA: 16

THO1: 7, 9.3

TPOX: 11

VK2/E6E7細(xì)胞Age: 32 years *****

Gender: female

Comments: The VK2/E6E7 (ATCC CRL-2616 cell line was established in 1996 from the normal vaginal mucosal tissue taken from a premenopausal woman undergoing anterior-posterior vaginal repair surgery. [52983]

The ectocervical Ect1/E6E7 (ATCC CRL-2614) and endocervical End1/E6E7 (ATCC CRL-2615) cell lines were established in 1996 from normal epithelial tissue taken from a premenopausal woman undergoing hysterectomy for endometriosis. [52983]

Cells at passage 3 were immortalized by transduction with the retroviral vector LXSN-16E6E7 in the presence of polybrene. Clones were selected in medium containing G418. [52983]

The endocervical cell line expresses characteristics of simple columnar epithelium, whereas the ectocervical and vaginal cell lines express characteristics of stratified squamous nonkeratinizing epithelia. [52984]

Without stimulation, all three cell lines produce macrophage colony-stimulating factor (M-CSF), transforming growth factor beta1, interleukin 8 (IL-8), prostaglandin E2, the secretory leukoproteinase inhibitor, and the polymeric immunoglobulin receptor. [52984]

The endocervical cell line (End1/E6E7), but not the others, also produce the lymphopoietic cytokines IL-6, IL-7, and consistently detectable levels of the chemokine known as "regulated-upon-activation, normal T cell expressed and secreted" (RANTES). [52984]

Stimulation with interferon gamma and tumor necrosis factor alpha (TNF alpha) induces or significantly up-regulates expression of several of the cytokines and chemokines as well as major histocompatibility complex (MHC) class II antigens in the lines. [52984]

Piliated, but not nonpiliated, Neisseria gonorrhoea strain F62 variants actively invade these epithelial cell lines.VK2/E6E7細(xì)胞 Invasion of these cells by green fluorescent protein-expressing gonococci is characterized by colocalization of gonococci with F actin. [53415]

These cell lines may provide the basis for valid, reproducible in vitro models for studies on cervicovaginal physiology and infections and for testing pharmacological agents for intravaginal application.

Propagation: ATCC complete growth medium: Keratinocyte-Serum Free medium (GIBCO-BRL 17005-042) with 0.1 ng/ml human recombinant EGF, 0.05 mg/ml bovine pituitary extract, and additional calcium chloride 44.1 mg/L (final concentration 0.4 mM)

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

Temperature: 37.0°C

Subculturing: Protocol: The cells should not be allowed to become confluent, subculture at 60 to 90% of confluence. Remove medium, and rinse with 0.25% trypsin, 0.53mM EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37C) until the cells detach.VK2/E6E7細(xì)胞 Neutralize the trypsin by adding a 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium containing 10% fetal bovine serum. Centrifuge the cell suspension at 1000 rpm for 10 minutes, resuspend the pellet in fresh serum-free growth medium, aspirate and dispense into new flasks. Cells will not attach well for 24 hours after subculture.

Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended

Medium Renewal: Every 2 to 3 days

Preservation: Freeze medium: A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium, 85%; fetal bovine serum, 10%; DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

Doubling Time: 24 hrs

References: 52983: Fichorova RN, et al. Generation of papillomavirus-immortalized cell lines from normal human ectocervical, endocervical, and vaginal epithelium that maintain expression of tissue-specific differentiation proteins. Biol. Reprod. 57: 847-855, 1999. PubMed: 9314589

52984: Fichorova RN, Anderson DJ. Differential expression of immunobiological mediators by immortalized human cervical and vaginal epithelial cells. Biol. Reprod. 60: 508-514, 1999. PubMed: 9916021

53415: Fichorova RN, et al. Distinct proinflammatory host responses to Neisseria gonorrhoeae infection in immortalized human cervical and vaginal epithelial cells. Infect. Immun. 69: 5840-5880, 2001. PubMed: 11500462

53416: Fichorova RN, et al. The molecular basis of nonoxynol-9-induced vaginal inflammation and its possible relevance to human immunodeficiency virus type 1 transmission. J. Infect. Dis. 184: 418-428, 2001. PubMed: 11471099