-
平臺編號：bio-68535
-
國際編號：CRL-11515?
-
細(xì)胞信息： 10.014 pRSV-T
-
規(guī)格：frozen
-
用途：ATCC原裝細(xì)胞
-
服務(wù)費(fèi)用：
加載中…… - 訂購
-
注意事項：僅用于科學(xué)研究或者工業(yè)應(yīng)用等非醫(yī)療目的不可用于人類或動物的臨床診斷或治療，非藥用，非食用（產(chǎn)品信息以出庫為準(zhǔn)）
10.014 pRSV-T
器官來源：眼
ATCC Number：CRL-11515?
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
運(yùn)輸方式：凍存運(yùn)輸
組織來源：cornea
生長狀態(tài)：貼壁生長
細(xì)胞形態(tài)：上皮樣
規(guī)格：0.5mg Designations: 10.014 pRSV-T
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
Permits/Forms: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.
DNA Profile (STR):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
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: Protocol:Subculture when 80% confluent
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. 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
ATCC Number：CRL-11515?
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
運(yùn)輸方式：凍存運(yùn)輸
組織來源：cornea
生長狀態(tài)：貼壁生長
細(xì)胞形態(tài)：上皮樣
規(guī)格：0.5mg Designations: 10.014 pRSV-T
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
Permits/Forms: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.
DNA Profile (STR):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
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: 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. 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
