2.040 pRSV-T细胞
运输方式： 冻存运输
细胞形态： 上皮样
细胞类型： 其他细胞类型
是否是肿瘤细胞： 0
物种来源： 人
组织来源： cornea
器官来源： 眼
生长状态： 贴壁生长
ATCC Number： CRL-11516™
2.040 pRSV-T细胞数量： 大量
Designations: 2.040 pRSV-T
Depositors: T Walker, Gillette Medical Evaluation Laboratories
Biosafety Level: 2 [Cells contain SV-40 and Rous Sarcoma viral sequences ]
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens
Morphology: epithelial
Source: Organ: eye
Tissue: cornea
Cell Type: 2.040 pRSV-T细胞epithelial transfected with a plasmid containing the SV40 early region
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Comments: A primary culture of normal corneal epithelium was immortalized by transfection with plasmid pRSV-T using lipofectamine overnight at 37C. pRSV-T contains the SV40 early region genes and the Rous Sarcoma virus long terminal repeat.
Propagation: ATCC complete growth medium: Keratinocyte-Serum Free medium (GIBCO-BRL 17005-042) with 5 ng/ml human recombinant EGF, 0.05 mg/ml bovine pituitary extract, 0.005 mg/ml insulin and 500 ng/ml hydrocortisone.
Temperature: 37.0°C
Subculturing: Protocol: The cells should be subcultured before reaching confluence.
Remove medium, add fresh 0.05% trypsin - 0.53 mM EDTA, rinse and remove trypsin. Allow the culture sit at room temperature (or 37C) until the cells detach. 2.040 pRSV-T细胞Neutralize the trypsin with fresh growth medium, centrifuge at 300 x g for 8 minutes, resuspend the pellet in fresh growth medium and dispense into new COATED flasks. Flasks used to propagate these cells must be coated (2 hours at 37C) with 0.01 mg/ml bovine serum albumin, 0.01 mg/ml fibronectin and 0.03 mg/ml bovine collagen type I .
Incubate at 37C with 5% CO2
Subcultivation Ratio: Subculture at 1:3 when 80% confluent.
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium, 82.5%; DMSO, 7.5%
Storage temperature: liquid nitrogen vapor phase
References: 34094: Walker TL, Kahn CR. 2.040 pRSV-T细胞Human corneal epithelial cell lines with extended lifespan. US Patent 5,672,498 dated Sep 30 1997
