A new rapid and non-radioactive assay for monitoring and determining the proliferation of retinal pigment epithelial cells.
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.
Yu
Hyeong Gon
HG
Department of Ophthalmology, Seoul National University College of Medicine, Seoul Artificial Eye Center, Seoul National University Hospital Clinical Research Institute, Nano Bioelectronics and Systems Research Center of Seoul National University, Korea.
Chung
Hum
H
Yu
Young Suk
YS
Seo
Jong-Mo
JM
Heo
Jang Won
JW
D002452:Cell Count; D002455:Cell Division; D002470:Cell Survival; D004396:Coloring Agents; D005434:Flow Cytometry; D005453:Fluorescence; D006801:Humans; D010078:Oxazines; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000502:physiology; D014343:Trypan Blue; D014966:Xanthenes
D004396:Coloring Agents; D010078:Oxazines; D014966:Xanthenes; C005843:resazurin; D014343:Trypan Blue