Purchase this article with an account.
T.R. Kinnick, B. McKay, J. Hu, D. Bok, A.D. Marmorstein; Differences in differentiation potential between human retinal pigment epithelial cell cultures derived from fetal and nonfetal donors . Invest. Ophthalmol. Vis. Sci. 2004;45(13):3672.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose:To characterize the differences between retinal pigment epithelium (RPE) cell cultures derived from donors ranging in age from 20 wks gestation through 76 years. Methods: RPE cells were harvested and maintained according to published methods (Hu and Bok, Mol Vis 7: 14–9, 2000; McKay, ARVO abstract #387, 2003) which incorporate a calcium switch. Pigmentation of cultures was observed and documented qualitatively. Transepithelial resistances (TERs) were measured directly from Millicell HA inserts (Millipore Corporation, Bedford, MA) using an EVOM Epithelial Voltohmmeter. Protein markers (CRALBP, bestrophin) of RPE differentiation were assessed by Western blot. Results: Using modified Chee's medium with low calcium, only cells derived from fetal donors exhibited satisfactory levels of proliferation without prior enzymatic dissociation. Cultures derived from donors of all ages could be induced to pigment via calcium switch. However, only cultures derived from fetal donors expressed bestrophin and exhibited a significant TER when cultured on porous supports. Conclusions: Our findings indicate that RPE pigmentation is stimulated by Ca–dependent cell adhesion, however, only RPE cells derived from fetal donors could be induced to exhibit properties of later stage RPE differentiation –– including a high TER and expression of bestrophin.
This PDF is available to Subscribers Only