ZFIN ID: ZDB-PUB-201202-18
Conditional reprograming culture conditions facilitate growth of lower grade glioma models
Yuan, M., White, D., Resar, L., Bar, E., Groves, M., Cohen, A., Jackson, E., Bynum, J., Rubens, J., Mumm, J., Chen, L., Jiang, L., Raabe, E., Rodriguez, F., Eberhart, C.G.
Date: 2020
Source: Neuro-Oncology   23(5): 770-782 (Journal)
Registered Authors: Mumm, Jeff
Keywords: BRAFV600E, Conditional reprogramming, NF1, Senescence, low grade glioma
MeSH Terms:
  • Animals
  • Astrocytoma*
  • Brain Neoplasms*
  • Cell Culture Techniques
  • Cellular Reprogramming*
  • Glioma*
  • Mice
  • Proto-Oncogene Proteins B-raf
  • Zebrafish
PubMed: 33258947 Full text @ Neuro Oncol.
The conditional reprogramming cell culture method was developed to facilitate growth of senescence-prone normal and neoplastic epithelial cells, and involves co-culture with irradiated fibroblasts and the addition of a small molecule Rho kinase (ROCK) inhibitor. The aim of this study was to determine whether this approach would facilitate the culture of compact low grade gliomas.
We attempted to culture 4 pilocytic astrocytomas, 2 gangliogliomas, 2 myxopapillary ependymomas, 2 anaplastic gliomas, 2 difficult-to-classify low grade neuroepithelial tumors, a desmoplastic infantile ganglioglioma, and an anaplastic pleomorphic xanthoastrocytoma using a modified conditional reprogramming cell culture approach.
Conditional reprogramming resulted in robust increases in growth for a majority of these tumors, with fibroblast conditioned media and ROCK inhibition both required. Switching cultures to standard serum containing media, or serum free neurosphere conditions, with or without ROCK inhibition, resulted in decreased proliferation and induction of senescence markers. ROCK inhibition and conditioned media both promoted Akt and Erk1/2 activation. Several cultures, including one derived from a NF1-associated pilocytic astrocytoma (JHH-NF1-PA1) and one from a BRAF p.V600E mutant anaplastic pleomorphic xanthoastrocytoma (JHH-PXA1), exhibited growth sufficient for preclinical testing in vitro. In addition, JHH-NF1-PA1 cells survived and migrated in larval zebrafish orthotopic xenografts, while JHH-PXA1 formed orthotopic xenografts in mice histopathologically similar to the tumor from which it was derived.
These studies highlight the potential for the conditional reprogramming cell culture method to promote the growth of glial and glioneuronal tumors in vitro, in some cases enabling the establishment of long-term culture and in vivo models.