Impact of Primary RPE Cells in a Porcine Organotypic Co-Cultivation Model
The pathological events of age-related macular degeneration are characterized by degenerative processes involving the photoreceptor cells, retinal pigment epithelium (RPE), and the Bruch’s membrane as well as choroidal alterations. To mimic in vivo interactions between photoreceptor cells and RPE ce...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-07-01
|
| Series: | Biomolecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2218-273X/12/7/990 |
| _version_ | 1797440594339954688 |
|---|---|
| author | Natalie Wagner Armin Safaei José Hurst Pia A. Vogt H. Burkhard Dick Stephanie C. Joachim Sven Schnichels |
| author_facet | Natalie Wagner Armin Safaei José Hurst Pia A. Vogt H. Burkhard Dick Stephanie C. Joachim Sven Schnichels |
| author_sort | Natalie Wagner |
| collection | DOAJ |
| description | The pathological events of age-related macular degeneration are characterized by degenerative processes involving the photoreceptor cells, retinal pigment epithelium (RPE), and the Bruch’s membrane as well as choroidal alterations. To mimic in vivo interactions between photoreceptor cells and RPE cells ex vivo, complex models are required. Hence, the aim of this study was to establish a porcine organotypic co-cultivation model and enlighten the interactions of photoreceptor and RPE cells, with a special emphasis on potential neuroprotective effects. Porcine neuroretina explants were cultured with primary porcine RPE cells (ppRPE) or medium derived from these cells (=conditioned medium). Neuroretina explants cultured alone served as controls. After eight days, RT-qPCR and immunohistology were performed to analyze photoreceptors, synapses, macroglia, microglia, complement factors, and pro-inflammatory cytokines (e.g., <i>IL1</i><i>B</i>, <i>IL6</i>, <i>TNF</i>) in the neuroretina samples. The presence of ppRPE cells preserved photoreceptors, whereas synaptical density was unaltered. Interestingly, on an immunohistological as well as on an mRNA level, microglia and complement factors were comparable in all groups. Increased <i>IL6</i> levels were noted in ppRPE and conditioned medium samples, while <i>TNF</i> was only upregulated in the ppRPE group. <i>IL1</i><i>B</i> was elevated in conditioned medium samples. In conclusion, a co-cultivation of ppRPE cells and neuroretina seem to have beneficial effects on the neuroretina, preserving photoreceptors and maintaining synaptic vesicles in vitro. This organotypic co-cultivation model can be used to investigate the complex interactions between the retina and RPE cells, gain further insight into neurodegenerative pathomechanisms occurring in retinal diseases, and evaluate potential therapeutics. |
| first_indexed | 2024-03-09T12:10:24Z |
| format | Article |
| id | doaj.art-2f421dac845d40dca794cc3d006ef47f |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-09T12:10:24Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-2f421dac845d40dca794cc3d006ef47f2023-11-30T22:52:35ZengMDPI AGBiomolecules2218-273X2022-07-0112799010.3390/biom12070990Impact of Primary RPE Cells in a Porcine Organotypic Co-Cultivation ModelNatalie Wagner0Armin Safaei1José Hurst2Pia A. Vogt3H. Burkhard Dick4Stephanie C. Joachim5Sven Schnichels6Experimental Eye Research Institute, University Eye Hospital, Ruhr-University Bochum, 44892 Bochum, GermanyExperimental Eye Research Institute, University Eye Hospital, Ruhr-University Bochum, 44892 Bochum, GermanyCentre for Ophthalmology, University Eye Hospital Tübingen, 72076 Tübingen, GermanyExperimental Eye Research Institute, University Eye Hospital, Ruhr-University Bochum, 44892 Bochum, GermanyExperimental Eye Research Institute, University Eye Hospital, Ruhr-University Bochum, 44892 Bochum, GermanyExperimental Eye Research Institute, University Eye Hospital, Ruhr-University Bochum, 44892 Bochum, GermanyCentre for Ophthalmology, University Eye Hospital Tübingen, 72076 Tübingen, GermanyThe pathological events of age-related macular degeneration are characterized by degenerative processes involving the photoreceptor cells, retinal pigment epithelium (RPE), and the Bruch’s membrane as well as choroidal alterations. To mimic in vivo interactions between photoreceptor cells and RPE cells ex vivo, complex models are required. Hence, the aim of this study was to establish a porcine organotypic co-cultivation model and enlighten the interactions of photoreceptor and RPE cells, with a special emphasis on potential neuroprotective effects. Porcine neuroretina explants were cultured with primary porcine RPE cells (ppRPE) or medium derived from these cells (=conditioned medium). Neuroretina explants cultured alone served as controls. After eight days, RT-qPCR and immunohistology were performed to analyze photoreceptors, synapses, macroglia, microglia, complement factors, and pro-inflammatory cytokines (e.g., <i>IL1</i><i>B</i>, <i>IL6</i>, <i>TNF</i>) in the neuroretina samples. The presence of ppRPE cells preserved photoreceptors, whereas synaptical density was unaltered. Interestingly, on an immunohistological as well as on an mRNA level, microglia and complement factors were comparable in all groups. Increased <i>IL6</i> levels were noted in ppRPE and conditioned medium samples, while <i>TNF</i> was only upregulated in the ppRPE group. <i>IL1</i><i>B</i> was elevated in conditioned medium samples. In conclusion, a co-cultivation of ppRPE cells and neuroretina seem to have beneficial effects on the neuroretina, preserving photoreceptors and maintaining synaptic vesicles in vitro. This organotypic co-cultivation model can be used to investigate the complex interactions between the retina and RPE cells, gain further insight into neurodegenerative pathomechanisms occurring in retinal diseases, and evaluate potential therapeutics.https://www.mdpi.com/2218-273X/12/7/990co-cultivationcomplement systemcytokinesporcineretinaretinal pigment epithelium (RPE) cells |
| spellingShingle | Natalie Wagner Armin Safaei José Hurst Pia A. Vogt H. Burkhard Dick Stephanie C. Joachim Sven Schnichels Impact of Primary RPE Cells in a Porcine Organotypic Co-Cultivation Model Biomolecules co-cultivation complement system cytokines porcine retina retinal pigment epithelium (RPE) cells |
| title | Impact of Primary RPE Cells in a Porcine Organotypic Co-Cultivation Model |
| title_full | Impact of Primary RPE Cells in a Porcine Organotypic Co-Cultivation Model |
| title_fullStr | Impact of Primary RPE Cells in a Porcine Organotypic Co-Cultivation Model |
| title_full_unstemmed | Impact of Primary RPE Cells in a Porcine Organotypic Co-Cultivation Model |
| title_short | Impact of Primary RPE Cells in a Porcine Organotypic Co-Cultivation Model |
| title_sort | impact of primary rpe cells in a porcine organotypic co cultivation model |
| topic | co-cultivation complement system cytokines porcine retina retinal pigment epithelium (RPE) cells |
| url | https://www.mdpi.com/2218-273X/12/7/990 |
| work_keys_str_mv | AT nataliewagner impactofprimaryrpecellsinaporcineorganotypiccocultivationmodel AT arminsafaei impactofprimaryrpecellsinaporcineorganotypiccocultivationmodel AT josehurst impactofprimaryrpecellsinaporcineorganotypiccocultivationmodel AT piaavogt impactofprimaryrpecellsinaporcineorganotypiccocultivationmodel AT hburkharddick impactofprimaryrpecellsinaporcineorganotypiccocultivationmodel AT stephaniecjoachim impactofprimaryrpecellsinaporcineorganotypiccocultivationmodel AT svenschnichels impactofprimaryrpecellsinaporcineorganotypiccocultivationmodel |