Purchase this article with an account.
J.D. Rios-Garcia, H. Tran, M. Shatos, H. Urashima, D.A. Dartt; Effect of OPC-12759 on the Proliferation of Rat Conjunctival Goblet Cells in Culture . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3786.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: To determine if OPC-12759, 2-(4chlorobenzoylamino)-3-[2(1H)-quinolinon-4yi]-propionic acid, increases proliferation of rat conjunctival goblet cells in primary culture. Methods: Goblet cells were isolated from rat conjunctiva and propagated in RPMI 1640 supplemented with 10% fetal bovine serum (FBS). Subconfluent cultured cells were serum starved for 48 hours and then incubated with or without increasing concentrations of OPC-12759 (10-12 to 10-8 M) for 1 to 7 days. 10% FBS was used as positive control. Cell proliferation was determined by colorimetric MTT assay or immunofluorescence microscopy using an antibody against Ki-67, which is expressed in proliferating cells. Mounting medium containing DAPI was used to identify cell nuclei. The number of Ki-67 positive cells and total cells were counted and proliferating cells were expressed as percentage of total. To determine if proliferating cells were goblet cells, cultured cells were double-labeled with the lectin Ulex europeus agglutinin I (UEA-I) and with antibody against Ki-67. Results: Using the MTT assay, OPC-12759 induced a concentration- and time-dependent increase in goblet cell proliferation. OPC-12759 significantly increased cell proliferation by 1.9 fold at day 1 and 1.3 fold at day 3 of incubation. The effect of OPC-12579 was not detected at day 5 and 7. FBS significantly increased cell proliferation by about 2 to 3 fold during the entire incubation time. As determined by the number of Ki-67 positive cells, OPC-12759 (10-11 M) increased goblet cell proliferation 5.0 fold at day 1, 3.4 fold at day 3, and 2.3 at day 5 compared to the control. FBS induced a 10-fold increase in goblet cell proliferation on days 1, 3 and 5. The colocalization of Ki-67 and UEA-I immunoreactivity indicated that the proliferating cells were goblet cells. Conclusions: OPC-12759 stimulates the proliferation of conjunctival goblet cells in primary culture.
This PDF is available to Subscribers Only