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| Home > Publications database > Generation of patient-derived pediatric pilocytic astrocytoma in-vitro models using SV40 large T: evaluation of a modeling workflow. > print |
| 001 | 285655 | ||
| 005 | 20240229155115.0 | ||
| 024 | 7 | _ | |a 10.1007/s11060-023-04500-6 |2 doi |
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| 037 | _ | _ | |a DKFZ-2023-02478 |
| 041 | _ | _ | |a English |
| 082 | _ | _ | |a 610 |
| 100 | 1 | _ | |a Selt, Florian |0 P:(DE-He78)a23e88cc676489fe05be8c178ceaf58e |b 0 |e First author |u dkfz |
| 245 | _ | _ | |a Generation of patient-derived pediatric pilocytic astrocytoma in-vitro models using SV40 large T: evaluation of a modeling workflow. |
| 260 | _ | _ | |a Dordrecht [u.a.] |c 2023 |b Springer Science + Business Media B.V |
| 336 | 7 | _ | |a article |2 DRIVER |
| 336 | 7 | _ | |a Output Types/Journal article |2 DataCite |
| 336 | 7 | _ | |a Journal Article |b journal |m journal |0 PUB:(DE-HGF)16 |s 1703857023_31546 |2 PUB:(DE-HGF) |
| 336 | 7 | _ | |a ARTICLE |2 BibTeX |
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| 336 | 7 | _ | |a Journal Article |0 0 |2 EndNote |
| 500 | _ | _ | |a #EA:B310#LA:B310# / 2023 Dec;165(3):467-478 |
| 520 | _ | _ | |a Although pediatric low-grade gliomas (pLGG) are the most common pediatric brain tumors, patient-derived cell lines reflecting pLGG biology in culture are scarce. This also applies to the most common pLGG subtype pilocytic astrocytoma (PA). Conventional cell culture approaches adapted from higher-grade tumors fail in PA due to oncogene-induced senescence (OIS) driving tumor cells into arrest. Here, we describe a PA modeling workflow using the Simian Virus large T antigen (SV40-TAg) to circumvent OIS.18 pLGG tissue samples (17 (94%) histological and/or molecular diagnosis PA) were mechanically dissociated. Tumor cell positive-selection using A2B5 was perfomed in 8/18 (44%) cases. All primary cell suspensions were seeded in Neural Stem Cell Medium (NSM) and Astrocyte Basal Medium (ABM). Resulting short-term cultures were infected with SV40-TAg lentivirus. Detection of tumor specific alterations (BRAF-duplication and BRAF V600E-mutation) by digital droplet PCR (ddPCR) at defined time points allowed for determination of tumor cell fraction (TCF) and evaluation of the workflow. DNA-methylation profiling and gene-panel sequencing were used for molecular profiling of primary samples.Primary cell suspensions had a mean TCF of 55% (+/- 23% (SD)). No sample in NSM (0/18) and ten samples in ABM (10/18) were successfully transduced. Three of these ten (30%) converted into long-term pLGG cell lines (TCF 100%), while TCF declined to 0% (outgrowth of microenvironmental cells) in 7/10 (70%) cultures. Young patient age was associated with successful model establishment.A subset of primary PA cultures can be converted into long-term cell lines using SV40-TAg depending on sample intrinsic (patient age) and extrinsic workflow-related (e.g. type of medium, successful transduction) parameters. Careful monitoring of sample-intrinsic and extrinsic factors optimizes the process. |
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| 650 | _ | 7 | |a Circumvention of OIS |2 Other |
| 650 | _ | 7 | |a In-vitro models |2 Other |
| 650 | _ | 7 | |a Inducible SV40 large T |2 Other |
| 650 | _ | 7 | |a Pediatric low-grade glioma cell lines |2 Other |
| 650 | _ | 7 | |a Pilocytic astrocytoma |2 Other |
| 700 | 1 | _ | |a El Damaty, Ahmed |b 1 |
| 700 | 1 | _ | |a Schuhmann, Martin U |b 2 |
| 700 | 1 | _ | |a Sigaud, Romain |0 P:(DE-He78)a5e60710c7515b3e1de74ced6928a9dd |b 3 |u dkfz |
| 700 | 1 | _ | |a Ecker, Jonas |0 P:(DE-He78)3de637452ba900e2bdd359b8f41953bf |b 4 |u dkfz |
| 700 | 1 | _ | |a Sievers, Philipp |0 P:(DE-He78)8aad075b17d93a5636a34942bdbd7ee6 |b 5 |u dkfz |
| 700 | 1 | _ | |a Kocher, Daniela |0 P:(DE-He78)4679059a19b1f3cd341bd27d0ddb7c45 |b 6 |u dkfz |
| 700 | 1 | _ | |a Herold-Mende, Christel |b 7 |
| 700 | 1 | _ | |a Oehme, Ina |0 P:(DE-He78)908367a659dea9e28dac34592b3c46e5 |b 8 |u dkfz |
| 700 | 1 | _ | |a von Deimling, Andreas |0 P:(DE-He78)a8a10626a848d31e70cfd96a133cc144 |b 9 |u dkfz |
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| 700 | 1 | _ | |a Jones, David |0 P:(DE-He78)551bb92841f634070997aa168d818492 |b 12 |u dkfz |
| 700 | 1 | _ | |a Witt, Olaf |0 P:(DE-He78)143af26de9d57bf624771616318aaf7c |b 13 |u dkfz |
| 700 | 1 | _ | |a Milde, Till |0 P:(DE-He78)0be2f86573954f87e97f8a4dbb05cb0f |b 14 |e Last author |u dkfz |
| 773 | _ | _ | |a 10.1007/s11060-023-04500-6 |0 PERI:(DE-600)2007293-4 |n 3 |p 467-478 |t Journal of neuro-oncology |v 165 |y 2023 |x 0167-594X |
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