Document Type

Article

Publication Date

7-15-2023

Keywords

JGM

JAX Source

Bioengineering (Basel). 2023;10(7):840

ISSN

2306-5354

PMID

37508868

DOI

https://doi.org/10.3390/bioengineering10070840

Grant

We acknowledge support from the Connecticut Children’s Medical Center (CCMC) Strate- gic Research Fund (to M.D.T-S.), UConn Health Stimulus Award (to M.D.T.-S.) and CCMC Surgical Research Seed Grant (to M.D.T-S.). Part of this study was funded by the American Brain Tumor Asso- ciation Discovery Grant from an Anonymous Family Foundation to M.D.T.-S. (DG1900023). M.D.T.-S. first started the study in UConn Health, continued after she moved to the Jackson Laboratory and finished the study after she left the Jackson Laboratory.

Abstract

Despite in vivo malignancy, ependymoma lacks cell culture models, thus limiting therapy development. Here, we used a tunable three-dimensional (3D) culture system to approximate the ependymoma microenvironment for recapitulating a patient's tumor in vitro. Our data showed that the inclusion of VEGF in serum-free, mixed neural and endothelial cell culture media supported the in vitro growth of all four ependymoma patient samples. The growth was driven by Nestin and Ki67 double-positive cells in a putative cancer stem cell niche, which was manifested as rosette-looking clusters in 2D and spheroids in 3D. The effects of extracellular matrix (ECM) such as collagen or Matrigel superseded that of the media conditions, with Matrigel resulting in the greater enrichment of Nestin-positive cells. When mixed with endothelial cells, the 3D co-culture models developed capillary networks resembling the in vivo ependymoma vasculature. The transcriptomic analysis of two patient cases demonstrated the separation of in vitro cultures by individual patients, with one patient's culture samples closely clustered with the primary tumor tissue. While VEGF was found to be necessary for preserving the transcriptomic features of in vitro cultures, the presence of endothelial cells shifted the gene's expression patterns, especially genes associated with ECM remodeling. The homeobox genes were mostly affected in the 3D in vitro models compared to the primary tumor tissue and between different 3D formats. These findings provide a basis for understanding the ependymoma microenvironment and enabling the further development of patient-derived in vitro ependymoma models for personalized medicine.

Comments

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).

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