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| 001 | 265755 | ||
| 005 | 20231120155400.0 | ||
| 024 | 7 | _ | |a 10.3389/fncel.2023.1166641 |2 doi |
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| 037 | _ | _ | |a DZNE-2023-01030 |
| 041 | _ | _ | |a English |
| 082 | _ | _ | |a 610 |
| 100 | 1 | _ | |a Carido, Madalena |b 0 |
| 245 | _ | _ | |a Reliability of human retina organoid generation from hiPSC-derived neuroepithelial cysts. |
| 260 | _ | _ | |a Lausanne |c 2023 |b Frontiers Research Foundation |
| 336 | 7 | _ | |a article |2 DRIVER |
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| 520 | _ | _ | |a The possible applications for human retinal organoids (HROs) derived from human induced pluripotent stem cells (hiPSC) rely on the robustness and transferability of the methodology for their generation. Standardized strategies and parameters to effectively assess, compare, and optimize organoid protocols are starting to be established, but are not yet complete. To advance this, we explored the efficiency and reliability of a differentiation method, called CYST protocol, that facilitates retina generation by forming neuroepithelial cysts from hiPSC clusters. Here, we tested seven different hiPSC lines which reproducibly generated HROs. Histological and ultrastructural analyses indicate that HRO differentiation and maturation are regulated. The different hiPSC lines appeared to be a larger source of variance than experimental rounds. Although previous reports have shown that HROs in several other protocols contain a rather low number of cones, HROs from the CYST protocol are consistently richer in cones and with a comparable ratio of cones, rods, and Müller glia. To provide further insight into HRO cell composition, we studied single cell RNA sequencing data and applied CaSTLe, a transfer learning approach. Additionally, we devised a potential strategy to systematically evaluate different organoid protocols side-by-side through parallel differentiation from the same hiPSC batches: In an explorative study, the CYST protocol was compared to a conceptually different protocol based on the formation of cell aggregates from single hiPSCs. Comparing four hiPSC lines showed that both protocols reproduced key characteristics of retinal epithelial structure and cell composition, but the CYST protocol provided a higher HRO yield. So far, our data suggest that CYST-derived HROs remained stable up to at least day 200, while single hiPSC-derived HROs showed spontaneous pathologic changes by day 200. Overall, our data provide insights into the efficiency, reproducibility, and stability of the CYST protocol for generating HROs, which will be useful for further optimizing organoid systems, as well as for basic and translational research applications. |
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| 650 | _ | 7 | |a gliosis |2 Other |
| 650 | _ | 7 | |a hiPSC |2 Other |
| 650 | _ | 7 | |a human |2 Other |
| 650 | _ | 7 | |a neurodevelopment |2 Other |
| 650 | _ | 7 | |a organoid |2 Other |
| 650 | _ | 7 | |a pathology |2 Other |
| 650 | _ | 7 | |a retina |2 Other |
| 650 | _ | 7 | |a stem cells |2 Other |
| 700 | 1 | _ | |a Völkner, Manuela |0 P:(DE-2719)2810390 |b 1 |e First author |u dzne |
| 700 | 1 | _ | |a Steinheuer, Lisa Maria |b 2 |
| 700 | 1 | _ | |a Wagner, Felix |0 P:(DE-2719)2811441 |b 3 |u dzne |
| 700 | 1 | _ | |a Kurth, Thomas |b 4 |
| 700 | 1 | _ | |a Dumler, Natalie |0 P:(DE-2719)9001836 |b 5 |u dzne |
| 700 | 1 | _ | |a Ulusoy, Selen |0 P:(DE-2719)9001420 |b 6 |u dzne |
| 700 | 1 | _ | |a Wieneke, Stephanie |0 P:(DE-2719)2812103 |b 7 |
| 700 | 1 | _ | |a Norniella, Anabel Villanueva |b 8 |
| 700 | 1 | _ | |a Golfieri, Cristina |0 P:(DE-2719)2810498 |b 9 |
| 700 | 1 | _ | |a Khattak, Shahryar |b 10 |
| 700 | 1 | _ | |a Schoenfelder, Bruno |0 P:(DE-2719)9001669 |b 11 |u dzne |
| 700 | 1 | _ | |a Scamozzi, Maria |0 P:(DE-2719)9002606 |b 12 |u dzne |
| 700 | 1 | _ | |a Zoschke, Katja |0 P:(DE-2719)2810521 |b 13 |u dzne |
| 700 | 1 | _ | |a Canzler, Sebastian |b 14 |
| 700 | 1 | _ | |a Hackermüller, Jörg |b 15 |
| 700 | 1 | _ | |a Ader, Marius |b 16 |
| 700 | 1 | _ | |a Karl, Mike O |0 P:(DE-2719)2000041 |b 17 |e Last author |u dzne |
| 773 | _ | _ | |a 10.3389/fncel.2023.1166641 |g Vol. 17, p. 1166641 |0 PERI:(DE-600)2452963-1 |p 1166641 |t Frontiers in cellular neuroscience |v 17 |y 2023 |x 1662-5102 |
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