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ARPE-19 (ATCC® CRL-2302™)

Organism: Homo sapiens, human  /  Cell Type: Epithelial  /  Tissue: eye; retinal pigmented epithelium; retina  /  Disease: normal

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Organism Homo sapiens, human
Tissue eye; retinal pigmented epithelium; retina
Cell Type Epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease normal
Age 19 years
Gender male
Applications
This cell line is a transfection host.
Storage Conditions liquid nitrogen vapor phase
Karyotype diploid
Derivation
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 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.

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.
Clinical Data
male
19 years
Antigen Expression
RPE-specific markers CRALBP and RPE-65
Genes Expressed
RPE-specific markers CRALBP and RPE-65
Comments
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.
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%.
Subculturing
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.05% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. 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.
  4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  5. 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.
  6. 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
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
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
Name of Depositor LM Hjelmeland
Passage History
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 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.
The cells were subjected to selective trypsinization for the first four passages to remove superficial cells before passaging the cuboidal basal layer.
Year of Origin 1986
References

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

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

Holtkamp GM, et al. Polarized secretion of IL-6 and IL-8 by human retinal pigment epithelial cells. Clin. Exp. Immunol. 112: 34-43, 1998. PubMed: 9566787

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

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

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

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

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

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

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

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

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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Product Sheet
Product Sheet
Certificate of Analysis
Certificate of Analysis
SDS
SDS
References

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

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

Holtkamp GM, et al. Polarized secretion of IL-6 and IL-8 by human retinal pigment epithelial cells. Clin. Exp. Immunol. 112: 34-43, 1998. PubMed: 9566787

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

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

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

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

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

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

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

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

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