A New Rapid and Non-radioactive Assay for Monitoring and Determining the Proliferation of Retinal Pigment Epithelial Cells.
- Author:
Hyeong Gon YU
1
;
Hum CHUNG
;
Young Suk YU
;
Jong Mo SEO
;
Jang Won HEO
Author Information
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords: absorbance; alamarBlue; fluorescence; proliferation; retinal pigment epithelial cells
- MeSH: 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
- From:Korean Journal of Ophthalmology 2003;17(1):29-34
- CountryRepublic of Korea
- Language:English
- 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.
