“…We show response patterns with fRPE, suggesting significance of our results. Contrary to a report with short-term PC aRPE [9], we observed no differences in morphology or TER at longterm PC between purely mechanical and enzyme-harvested cultures (data not shown). We caution the interpretation of control cultures from younger and fetal donors: It is possible that a larger population of these donors may have altered the data obtained.…”
Dear Editor, Visual rehabilitation achieved with peripheral RPE autografts in exudative AMD is only modest. Moreover, complication rates using the popular RPE-choroid patch technique remain relatively high.Tissue-engineered transplants might improve the outcome of RPE transplantation. A culturing step could place cells on a substrate prior to transplantation, thereby optimizing delivery and long-term function [1]. This would require sufficient cellular plasticity (i.e., response to environmental stimuli) to achieve a well-differentiated, functional graft. While culture plasticity was confirmed for fetal cells or RPE cell lines [2,3], only sparse information is available for aged RPE [4].We studied the influence of culture conditions on select differentiation characteristics in long-term primary RPE cultures from ten human donors above age 55, and compared them to a 23-year-old and two fetal donors (20 and 21 weeks of gestation respectively). The main outcome measures at 6 weeks post-confluence (PC) were morphology and transepithelial resistance (TER), an electrophysiological method for assessing tight junctions build-up.Eyes were preserved within 6 hours of death. Adult primary RPE cultures (aRPE) were initiated from: (1) mechanically scraped RPE, or (2) collagenase pretreated (1 mg/ml for 40 minutes, CLS
“…We show response patterns with fRPE, suggesting significance of our results. Contrary to a report with short-term PC aRPE [9], we observed no differences in morphology or TER at longterm PC between purely mechanical and enzyme-harvested cultures (data not shown). We caution the interpretation of control cultures from younger and fetal donors: It is possible that a larger population of these donors may have altered the data obtained.…”
Dear Editor, Visual rehabilitation achieved with peripheral RPE autografts in exudative AMD is only modest. Moreover, complication rates using the popular RPE-choroid patch technique remain relatively high.Tissue-engineered transplants might improve the outcome of RPE transplantation. A culturing step could place cells on a substrate prior to transplantation, thereby optimizing delivery and long-term function [1]. This would require sufficient cellular plasticity (i.e., response to environmental stimuli) to achieve a well-differentiated, functional graft. While culture plasticity was confirmed for fetal cells or RPE cell lines [2,3], only sparse information is available for aged RPE [4].We studied the influence of culture conditions on select differentiation characteristics in long-term primary RPE cultures from ten human donors above age 55, and compared them to a 23-year-old and two fetal donors (20 and 21 weeks of gestation respectively). The main outcome measures at 6 weeks post-confluence (PC) were morphology and transepithelial resistance (TER), an electrophysiological method for assessing tight junctions build-up.Eyes were preserved within 6 hours of death. Adult primary RPE cultures (aRPE) were initiated from: (1) mechanically scraped RPE, or (2) collagenase pretreated (1 mg/ml for 40 minutes, CLS
“…Although transplanted autologous IPE cells in vivo phagocytose photoreceptor outer segments in the subretinal space (Thumann et al 1999a), little is known of the retinoid metabolism of transplanted IPE cells in the subretinal space. Iris pigment epithelial cells in culture did not show any retinoid metabolism (Von Recum et al 1999), but the presence of mRNA for cellular retinaldehyde binding protein (CRALBP) and for some related substances suggests the possibility that IPE cells may be able to metabolize retinol in a proper micro-environment (Thumann et al 1999a). The transdifferentiation ability and lens-forming potency of IPE cells has been reported previously (Eguchi & Shingai 1971) and has been recently documented in chicken IPE in EdFPH media (Kosaka et al 1998).…”
ABSTRACT.Purpose: To investigate photoreceptor survival in transplantation of non-cultured iris pigment epithelial (IPE) cells to the subretinal space in a prospective experimental study. Methods: Upper iridectomies were carried out in the right eyes of 37 pigmented rabbits. Suspensions of freshly harvested autologous IPE cells (without culturing) were prepared and injected into the subretinal space of the same eye. Follow-up examinations were carried out using ophthalmoscopy and colour fundus photography. The rabbits were killed at 1, 2, 3 and 6 months, respectively, and the eyes examined with light and electron microscopy. Results: On histological examination, the photoreceptor cells were found to be well-preserved in grafted areas at 1-3 months. At 6 months, the photoreceptors generally disclosed a normal nuclear layer and long outer segments when overlying areas with single cells or clusters of transplanted IPE cells. Multilayers of cells in abundance, including native RPE cells and macrophages (stained with RAM 11), particularly under microfolds of the neural retina, were occasionally associated with photoreceptor damage and nuclear drop out from the outer retinal layer. There was no inflammatory response in the choroid and the choriocapillaris remained patent.
Conclusion:The experiments show that grafting freshly harvested autologous IPE cells to the subretinal space is feasible and that the photoreceptors generally survive for at least 6 months when overlying the transplanted areas. Multilayers of abundant cells in the subretinal space may induce adverse focal effects on adjacent photoreceptors.
“…The surface is irradiated with UV light to cross-link the copolymer through dimerization of the cinnamoyl groups. 25 Similar to plasma polymerized pNIPAAm, the thickness of the film does not affect cell attachment and proliferation.…”
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