Establishing Neural Stem Cell and Medulloblastoma Cell Lines

DKFZ-EP1NS [25 (link)], SJ-BT57 [52 (link)], and human fetal neural stem cells obtained from C. Watts, University of Cambridge were cultivated as neural stem cells using neurosphere medium (Invitrogen, Karlsruhe, Germany) with growth factor supplements as described [25 (link)] Medulloblastoma cells DAOY (obtained from American Type Culture Collection) and UW228-3 cells (obtained from Steven Clifford, Newcastle, United Kingdom), were cultivated adherently using normal DMEM (Dubecco's modified Eagle medium, Invitrogen, Karlsruhe, Germany). Based on DNA methylation profiling using Illumina 450k methylation array [11 (link)], cell lines EP1NS and SJ-BT57 were classified as preclinical RELA-positive (EP1NS) and posterior fossa Group A (SJ-BT57) models. Additionally, according to the study of Parker and colleagues [15 (link)], the RELA-C11orf95 fusions was detected in EP1NS by reverse-transcription PCR, detailed description can be found elsewhere [11 (link), 15 (link)].

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Tzaridis T., Milde T., Pajtler K.W., Bender S., Jones D.T., Müller S., Wittmann A., Schlotter M., Kulozik A.E., Lichter P., Collins V.P., Witt O., Kool M., Korshunov A., Pfister S.M, & Witt H. (2016). Low-dose Actinomycin-D treatment re-establishes the tumoursuppressive function of P53 in RELA-positive ependymoma. Oncotarget, 7(38), 61860-61873.

Publication 2016

Dubecco's modified Eagle medium 450k methylation array

Cell lines Cells Eagle Fetal Growth factor Human Medulloblastoma Methylation Neural stem cells Rela Reverse transcription Supplements

Corresponding Organization :

Other organizations : German Cancer Research Center, Heidelberg University, National Center for Tumor Diseases, University Hospital Heidelberg, University of Cambridge

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Variable analysis

independent variables

Cell lines used: DKFZ-EP1NS, SJ-BT57, and human fetal neural stem cells
Cultivation conditions: neurosphere medium with growth factor supplements, and normal DMEM medium

dependent variables

DNA methylation profiling using Illumina 450k methylation array
Detection of RELA-C11orf95 fusion by reverse-transcription PCR

control variables

Not explicitly mentioned

controls

Positive control: Not mentioned
Negative control: Not mentioned

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