Korean J Ophthalmol.  2003 Jun;17(1):29-34. 10.3341/kjo.2003.17.1.29.

A New Rapid and Non-radioactive Assay for Monitoring and Determining the Proliferation of Retinal Pigment Epithelial Cells

• Yu HG
• Chung H
• Yu YS
• Seo JM
• Heo JW

• KMID: 754339
• DOI: http://doi.org/10.3341/kjo.2003.17.1.29

Abstract

AlamarBlue is used to induce color and fluorescence in the microenvironment of activated cells. The alamarBlue assay was studied to determine if it could be used as a method of evaluating the number of retinal pigment epithelial (RPE) cells. A series of two-fold dilutions of RPE cells were placed into 96-well culture plates. The alamarBlue was added to the culture media after attaching the cells. The absorbance and fluorescence were measured consecutively at various intervals over a period of 24 hr. Cell viability were evaluated by means of the trypan blue exclusion method and flow cytometry using a combination of propidium iodide and annexin V was done to prove the safety of alamarBlue assay to the cells. Both the absorbance and the fluorescence had a linear relationship with the number of RPE cells. Exposing the RPE cells to alamarBlue was not detrimental to the cells. In conclusion, the alamarBlue assay constitutes a one-step, extremely simple, reproducible, economical and non-toxic procedure for evaluating the number of viable RPE cells.

Keyword

absorbance; alamarBlue; fluorescence; proliferation; retinal pigment epithelial cells

MeSH Terms

Cell Count
Cell Division
Cell Survival
*Dyes
Flow Cytometry
Fluorescence
Human
Pigment Epithelium of Eye/*cytology/physiology
Support, Non-U.S. Gov't
Trypan Blue