10.014 pRSV-T细胞
器官来源： 眼
ATCC Number： CRL-11515™
数量： 大量
细胞类型： 其他细胞类型
是否是肿瘤细胞： 0
物种来源： 人
运输方式： 冻存运输
组织来源： cornea
生长状态： 贴壁生长
细胞形态： 上皮样
Designations: 10.014 pRSV-T
10.014 pRSV-T细胞Depositors: CR Kahn, 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: epithelial transfected with a plasmid containing the SV40 early region
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DNA Profile (STR): Amelogenin: X,Y
CSF1PO: 10
D13S317: 12,13
D16S539: 8,13
D5S818: 12,13
D7S820: 8,10
10.014 pRSV-T细胞THO1: 6,8
TPOX: 8,10
vWA: 15
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
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Subculturing: 10.014 pRSV-T细胞Protocol:Subculture when 80% confluent
remove medium
briefly rinse the cell layer with 0.05% (w/v) Trypsin-53mM EDTA solution
add Trypsin-EDTA solution to flask and incubate at 37�C for up to three minutes to facilitate dispersal. Observe cells under an inverted microscope. Hit or shake the flask if some cells remain attached
neutralize the trypsin with 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 (for at least 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. Cells will not be well attached for the first 24 hours after subculture.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended when 80% confluent
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: complete growth medium supplemented with 7.5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
References: 34094: Walker TL, Kahn CR. 10.014 pRSV-T细胞Human corneal epithelial cell lines with extended lifespan. US Patent 5,672,498 dated Sep 30 1997
46895: 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
