ARPE-19 are human-derived spontaneously arisen retinal pigment epithelial cells. They are used to study various aspects of retinal biology, pathological conditions, and therapeutic interventions (pharmacology). This article aims to provide comprehensive insights into ARPE-19 immortalized cells. Mainly, general characteristics, culture conditions, and diverse research applications of the cell line will be discussed. Therefore, by reading this, you’ll gain a thorough understanding of:

  1. ARPE-19 cells: Origin and general attributes
  2. ARPE-19 cell line: Culturing information
  3. Advantages & Limitations of ARPE-19 cells
  4. Applications of ARPE-19 cell line in research
  5. ARPE-19 cells: Research publications
  6. Resources for ARPE-19 cell line: Protocols, Videos, and More

1.      ARPE-19 cells: Origin and general attributes

Knowing a cell line’s origin and general attributes is imperative for its effective use in research. This article section will cover all the information about the ARPE cell line. Such as, What is the ARPE-19 cell line? Why use ARPE-19 cells? What is the ARPE-19/HPV-16 cell line? Are ARPE-19 cells immortal? What is ARPE 19 cell morphology and size?

  • Immortalized retinal pigment epithelial cells line, ARPE-19, was derived from a 19-year-old man’s eyes who died from head trauma in an accident. It was established by Amy Aotaki-Keen in 1986.
  • These cells express retinal pigment epithelial cell markers, i.e., CRALBP and RPE-65, suggesting they can make stable monolayers characterized by morphological and functional polarity.
  • ARPE-19 cells possess an epithelial cell-like morphology.
  • ARPE-19 cells mostly have a normal karyotype except for one deletion and addition in the long arm of chromosome 9 and 19, respectively. Moreover, some aneuploidies are also observed [1].

Scanning electron micrograph (SEM) of retina rods and cones.

2.      ARPE-19 cell line: Culturing information

Basic cell culture information is crucial for properly handling and maintaining a cell line. This section will help you learn the key points for the ARPE-19 cell line culturing. You will know: what is arpe-19 doubling time? What is the ARPE-19 seeding density? What is the ARPE-19 cell density? What is ARPE-19 freezing media? How do you culture the ARPE-19 cell line?

Key Points for Culturing ARPE-19 Cells

Population Doubling Time:

The ARPE-19 doubling time is approximately 55- 65 hours. They may undergo up to 48 population doublings.

Adherent or in Suspension:

ARPE-19 is an adherent cell line.

Sub-cultivation ratio:

The sub-cultivation ratio of ARPE-19 is 1:3 to 1:5. Adherent cells are rinsed with 1x PBS and incubated with dissociation solution, accutase, for 8 to 10 min. Detached cells are added with fresh medium and centrifuged. The cell pellet was again resuspended and poured into a culture flask containing fresh medium.

Growth Medium:

DMEM or Ham’s F12 medium is used to culture the ARPE-19 cell line. The ARPE-19 medium is supplemented with 5% FBS, 3.1 g/L Glucose, 15 mM HEPES, 1.6 mM L-Glutamine, 1.0 mM Sodium pyruvate, and 1.2 g/L NaHCO3. Media is replaced 2 to 3 times per week.

Growth Conditions:

ARPE-19 cells are kept in a humidified incubator at 37°C temperature with a 5% CO2 supply.


The cell line can be stored in the vapour phase of liquid nitrogen or at below -150°C temperature to protect the viability of cells for the long term.

Freezing Process and Medium:

CM-1 or CM-ACF are used as ARPE-19 freezing media freezing media. Briefly, cells are frozen through a slow freezing method that permits only 1 °C per minute decrease in temperature and protects cells from shock.

Thawing Process:

Cells are thawed in a water bath pre-set at 37°C. After a small ice clump is left, cells are added to fresh culture medium and centrifuged. This removed freezing media elements. Next, the cell pellet is resuspended, and cells are dispensed into a flask for culturing.

Biosafety Level:

ARPE-19 cells are handled in biosafety 1 laboratories.


ARPE-19 cells at 10x magnification post-subcultivation and semi-confluent.

3.      Advantages & Limitations of ARPE-19 cells

ARPE-19 cells are widely used in retinal cell biology studies. Like other cells, they are also associated with some advantages and limitations. A few of them are listed in this section:


The main advantages of the ARPE-19 cell line include:

Retinal cell model

ARPE-19 cells closely resemble human retinal pigment epithelial cells, making them ideal for studying retinal diseases and drug testing.

Stable growth rate

These cells exhibit stable growth and can be maintained over extended periods, facilitating long-term experiments.

Transfection amenability

The ARPE-19 cell line is an excellent transfection host, widely used in both transient and stable expression studies.



Here are some limitations associated with the ARPE-19 cell line:

Limited Differentiation

ARPE-19 differentiation is limited compared to primary retinal cells. This may potentially impact certain differentiation-related studies.


4.      Applications of ARPE-19 cell line in research

The ARPE-19 cell line has numerous applications in retinal research. Here, we have talked about some specific and significant research uses of this retinal pigment epithelial cell line.

  • Retinal disease research: ARPE-19 cells offer valuable insights into retinal pathogenesis. Researchers use cells to investigate disease mechanisms and potential treatments. A study conducted in 2020 found that circular RNA hsa_circ_0041795 interacts with miRNA-646 and VEGFC to facilitate high glucose-induced injury in human retinal pigment epithelial cells ARPE-19. Therefore, the study proposes this circular RNA as an effective therapeutic and diagnostic target to combat diabetic retinopathy [2]. Similarly, Jing Yang and colleagues used ARPE-19 cells and provided insights into the pathogenesis of diabetic retinopathy. They studied that inhibiting lncRNA SNHG1 (Small Nucleolar RNA Host Gene 1) can suppress inflammatory response and epithelial to mesenchymal transition of high glucose-treated ARPE-19 cells [3].
  • Drug testing: ARPE-19 cells are used to evaluate the efficacy and safety of drugs and compounds, helping develop treatments or therapies for retinal diseases. For instance, a study carried out in 2019 found the protective effects of Syzygium malaccense bioactives against hydrogen peroxide-induced stress in human retinal pigment epithelial cells, ARPE-19 [4]. Following this, a study found the therapeutic role of Prunella vulgaris var. L extract against blue light-induced injury in ARPE-19 cells and mice model [5].

5.      ARPE-19 cells: Research publications

The following are some interesting research publications featuring the ARPE-19 retinal pigment epithelial cells.

Evaluation of microRNA responses in ARPE-19 cells against the oxidative stress

This research article was published in 2018 in the Cutaneous and Ocular Toxicology. This study evaluated the miRNA expression in response to oxidative stress induced in ARPE-19 human retinal pigment epithelial cells via hydrogen peroxide treatment.

ARPE-19 conditioned medium promotes neural differentiation of adipose-derived mesenchymal stem cells

This publication in the World Journal of Stem Cells (2021) proposed that ARPE-19 cells conditioned media contains growth factors that encourage the neural differentiation of mesenchymal stem cells derived from adipose tissues.

Quercetin inhibits the production of IL-1β-induced inflammatory cytokines and chemokines in ARPE-19 cells via the MAPK and NF-κB signaling pathways

This study was published in the International Journal of Molecular Sciences (2019). It states that quercetin protects IL-1β-stimulated release of chemokines in ARPE-19 cells by preventing the activation of MAPK and NF-κB cascades to improve the inflammatory response.

Resveratrol modulates SIRT1 and DNMT functions and restores LINE-1 methylation levels in ARPE-19 cells under oxidative stress and inflammation

This research article was published in the International Journal of Molecular Sciences (2018). This study evaluated the possible effects of oxidative stress and inflammation on SIRT1 (Sirtuin 1) and DNMTs (DNA methyltransferases) functions as well as on LINE-1 (long interspersed nuclear element-1) methylation in ARPE-19 cells.

Protective Effect of Chrysanthemum boreale Flower Extracts against A2E-Induced Retinal Damage in ARPE-19 Cell

This article in Antioxidants (2022) found that Chrysanthemum boreale Flower Extracts exert protective effects against N-retinylidene-N-retinylethanolamine (A2E) induced retinal damage in ARPE-19 cells.

6.      Resources for ARPE-19 cell line: Protocols, Videos, and More

ARPE-19 is a widely used retinal epithelial cell line. The available resources covering ARPE-19 cell culturing and transfection protocols are listed here:

  • ARPE-19 transfection: This video is a step-by-step guide to learning transfection protocol for the ARPE-19 cell line.

Here are some resources describing the ARPE-19 cell culture protocol:

  • ARPE-19 cell culture protocol: This link contains information about ARPE-19 cell culturing and maintenance. It comprises information about the ARPE-19 medium, culturing conditions, protocols for subculturing, and handling proliferative and cryopreserved cultures.


  1. Schnichels, S., et al., Retina in a dish: Cell cultures, retinal explants and animal models for common diseases of the retina. Progress in retinal and eye research, 2021. 81: p. 100880.
  2. Sun, H. and X. Kang, hsa_circ_0041795 contributes to human retinal pigment epithelial cells (ARPE 19) injury induced by high glucose via sponging miR-646 and activating VEGFC. Gene, 2020. 747: p. 144654.
  3. Yang, J., et al., Silenced SNHG1 Inhibited Epithelial-Mesenchymal Transition and Inflammatory Response of ARPE-19 Cells Induced by High Glucose. J Inflamm Res, 2021. 14: p. 1563-1573.
  4. Arumugam, B., et al., Protective effect of myricetin derivatives from Syzygium malaccense against hydrogen peroxide-induced stress in ARPE-19 cells. Molecular vision, 2019. 25: p. 47.
  5. Kim, J., K. Cho, and S.-Y. Choung, Protective effect of Prunella vulgaris var. L extract against blue light induced damages in ARPE-19 cells and mouse retina. Free Radical Biology and Medicine, 2020. 152: p. 622-631.