2.040 pRSV-T細(xì)胞

運(yùn)輸方式： 凍存運(yùn)輸

細(xì)胞形態(tài)： 上皮樣

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

組織來源： cornea

器官來源： 眼

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

ATCC Number： CRL-11516?

2.040 pRSV-T細(xì)胞數(shù)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細(xì)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細(xì)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細(xì)胞Human corneal epithelial cell lines with extended lifespan. US Patent 5,672,498 dated Sep 30 1997