ARPE-19細(xì)胞

運(yùn)輸方式： 凍存運(yùn)輸

器官來(lái)源： 眼

組織來(lái)源： retinal pigmented epithelium; retina

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

細(xì)胞形態(tài)： 上皮樣

ATCC Number： CRL-2302?

數(shù)量： 大量

相關(guān)**： 正常

年限： 19 years

是否是腫瘤細(xì)胞： 0

物種來(lái)源： 人

Designations: ARPE-19

Depositors: LM Hjelmeland

ARPE-19細(xì)胞Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: epithelial

Source: Organ: eye

Tissue: retinal pigmented epithelium; retina

Disease: normal

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Applications: transfection host

Antigen Expression: RPE-specific markers CRALBP and RPE-65

DNA Profile (STR): Amelogenin: X,Y

CSF1PO: 11

D13S317: 11,12

D16S539: 9,11

D5S818: 13

D7S820: 9,11

THO1: 6,9.3

TPOX: 9,11

vWA: 16,19

Cytogenetic Analysis: diploid

Age: 19 years

ARPE-19細(xì)胞Gender: male

Comments: ARPE-19 is a spontaneously arising retinal pigment epithelia (RPE) cell line derived in 1986 by Amy Aotaki-Keen from the normal eyes of a 19-year-old male who died from head trauma in a motor vehicle accident.

The cells were subjected to selective trypsinization for the first four passages to remove superficial cells before passaging the cuboidal basal layer.

By passage 5, the cultures appeared to be rapidly growing RPE cells, which would form cobblestone monolayers, which pigmented after several months in culture.

The line was established in a 1:1 mixture of Dulbecco's modified Eagles medium and Ham's F12 medium with HEPES buffer containing 20% fetal bovine serum, 56 mM final concentration sodium bicarbonate and 2 mM L-glutamine and incubated at 37C in 10% CO2.

These cells form stable monolayers, which exhibit morphological and functional polarity. ARPE-19 expresses the RPE-specific markers CRALBP and RPE-65.

The cells exhibit morphological polarization when plated on laminin-coated Transwell-COL filters in medium with a low serum concentration.

They form tight-junctions with transepithelial resistance of monolayers reaching a maximum of 50 to 100 ohms/cm2 after 4 weeks of culture.

The cells are diploid and can be carried for over 30 passages. Progeny were found to undergo an additional 48 population doublings in longevity trials performed during characterization at ATCC .

Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No. 30-2006. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37.0°C

Subculturing: Protocol:

Remove and discard culture medium.

Briefly rinse the cell layer with 0.05% (w/v)ARPE-19細(xì)胞 Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.

Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).

Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37?C to facilitate dispersal.

Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.

To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes.Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.

Incubate cultures at 37?C.

Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended

Medium Renewal: Two to three times weekly

Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2006

recommended serum:ATCC 30-2020

ATCC CRL-2502 is derived from ATCC CRL-2302

References: 34497: Dunn KC, et al. ARPE-19, A human retinal pigment epithelial cell line with differentiated properties. Exp. Eye Res. 62: 155-169, 1996. PubMed: 8698076

48288: Maidji E, et al. Accessory human cytomegalovirus glycoprotein US9 in the unique short component of the viral genome promotes cell-to-cell transmission of virus in polarized epithelial cells. J. Virol. 70: 8402-8410, 1996. PubMed: 8970961

48289: Holtkamp GM, et al. ARPE-19細(xì)胞Polarized secretion of IL-6 and IL-8 by human retinal pigment epithelial cells. Clin. Exp. Immunol. 112: 34-43, 1998. PubMed: 9566787

48291: Finnemann SC, et al. Phagocytosis of rod outer segments by retinal pigment epithelial cells requires alpha(v)beta5 integrin for binding but not for internalization. Proc. Natl. Acad. Sci. USA 94: 12932-12937, 1997. PubMed: 9371778

48292: Handa JT, et al. The advanced glycation endproduct pentosidine induces the expression of PDGF-B in human retinal pigment epithelial cells. Exp. Eye Res. 66: 411-419, 1998. PubMed: 9593635

48293: Dunn KC, et al. Use of the ARPE-19 cell line as a model of RPE polarity: basolateral secretion of FGF5.. Invest. Ophthalmol. Vis. Sci. 39: 2744-2749, 1998. PubMed: 9856785

48294: Tugizov S, et al. An acidic cluster in the cytosolic domain of human cytomegalovirus glycoprotein B is a signal for endocytosis from the plasma membrane. J. Virol. 73: 8677-8688, 1999. PubMed: 10482621

48295: Orten DJ, et al. Analysis of DNA elements that modulate myosin VIIA expression in humans. Hum. Mutat. 14: 354, 1999. PubMed: 10502787

48296: Rajan PD, et al. Expression of the extraneuronal monoamine transporter in RPE and neural retina. Curr. Eye Res. 20: 195-204, 2000. PubMed: 10694895

48297: Janssen JJ, et al. Retinoic acid delays transcription of human retinal pigment neuroepithelium marker genes in ARPE-19 cells. Neuroreport 11: 1571-1579, 2000. PubMed: 10841379

48298: Udono T, et al. Adrenomedullin in cultured human retinal pigment epithelial cells. Invest. Ophthalmol. Vis. Sci. 41: 1962-1970, 2000. PubMed: 10845623

48299: Sparrow JR, et al. ARPE-19細(xì)胞The lipofuscin fluorophore A2E mediates blue light-induced damage to retinal pigmented epithelial cells. Invest. Ophthalmol. Vis. Sci. 41: 1981-1989, 2000. PubMed: 10845625