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微信小章| 产品名称： | 10.014 pRSV-T |
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| 商品货号： | TS215236 |
| Organism： | Homo sapiens, human |
| Tissue： | eye, cornea |
| Cell Type： | epithelial cells transfected with a plasmid containing the SV40 early region |
| Product Format： | frozen |
| Morphology： | epithelial |
| Culture Properties： | adherent |
| Biosafety Level： | 2 Cells contain SV-40 and Rous Sarcoma viral 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. |
| Storage Conditions： | liquid nitrogen vapor phase |
| Disclosure： | This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC. |
| Images： |  |
| Derivation： | A primary culture of normal corneal epithelium was immortalized by transfection with plasmid pRSV-T using lipofectamine overnight at 37°C. pRSV-T contains the SV40 early region genes and the Rous Sarcoma virus long terminal repeat. |
| Comments： | On a personal communication with the ATCC, the depositor commented that these cells have a finite life span.
Depositor Reference: Walker TL, Kahn CR. Human corneal epithelial cell lines with extended lifespan. US Patent 5,672,498. |
| Complete Growth Medium： | The base medium for this cell line is Keratinocyte-Serum Free Medium (ThermoFisher cat # 17005-042) with 5 ng/ml human recombinant EGF, 0.05 mg/ml bovine pituitary extract. To make the complete medium add the following components to the base medium: 0.005 mg/ml human recombinant insulin (ThermoFisher cat # 12585-014) and 500 ng/ml hydrocortisone (Sigma cat # H0135). |
| 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. Note: The flasks used should be precoated with a mixture of 0.01 mg/mL fibronectin, 0.03 mg/mL bovine collagen type I and 0.01 mg/mL bovine serum albumin. Subculture when 80% confluent.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended when 80% confluent 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.xa0xa0 |
| Cryopreservation： | Freeze medium: complete growth medium supplemented with 7.5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions： | Temperature: 37°C Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| STR Profile： | Amelogenin: X,Y CSF1PO: 10 D13S317: 12,13 D16S539: 8,13 D5S818: 12,13 D7S820: 8,10 THO1: 6,8 TPOX: 8,10 vWA: 15 |
| Name of Depositor： | CR Kahn, Gillette Medical Evaluation Laboratories |
| U.S. Patent Number： | |
| References： | Walker TL, Kahn CR. Human corneal epithelial cell lines with extended lifespan. US Patent 5,672,498 dated Sep 30 1997 Kruszewski FH, et al. Evaluation of a human corneal epithelial cell line as an in vitro model for assessing ocular irritation. Fundam. Appl. Toxicol. 36: 130-140, 1997. PubMed: 9143482 |