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.

This cell line is a transfection host.

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.

male

19 years

RPE-specific markers CRALBP and RPE-65

RPE-specific markers CRALBP and RPE-65

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.

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¡ãC 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¡ãC.

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

Medium Renewal: Two to three times weekly

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

Storage temperature: liquid nitrogen vapor phase

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

Temperature: 37¡ãC