Characterization of Native Extracellular Matrix of Patient-Derived Glioblastoma Multiforme Organoids.

in Tissue engineering. Part A by Alexandra D Avera, Daniel J Gibson, Macy L Birge, Taylor N Schnorbus, Isabella M Concannon, Yonghyun Kim

TLDR

  • Too Long; Didn't Read: This study developed a method to generate glioblastoma multiforme (GBM) organoids (GBOs) without exogenous extracellular matrices (ECMs) and without cell culture media changes, producing stable tissue-like GBOs with natively produced ECMs.
  • Key Insights: The study observed a transition from homogenous cell populations to tissue-like structures, with the greatest gene expression changes occurring when GBOs reached 2 mm in diameter, and native ECM synthesis was confirmed through quantitative and histochemical assessments.

Abstract

Model systems play a crucial role in biological and biomedical research, especially in the search for new treatments for challenging diseases such as glioblastoma multiforme (GBM). Organoids are 3Dmulticellular "middle-ground" model systems that recapitulate highly organized and heterogeneousorgan-like systems, often through stem cell differentiation. Incorporating Matrigel™ or other exogenous extracellular matrices (ECMs) that do not naturally occur in the human body is common practice for organoid generation, ignoring the role of dynamic reciprocity between the cells and the ECM in tissue development. In this study, we describe a method to develop GBM organoids (GBOs) from cells without the need for exogenous ECM encapsulation and without cell culture media changes to produce stable tissue-like organoids that reach a 4 mm diameter in as little as 6 weeks. We observed a transition from homogenous cell populations to tissue-like structures when GBOs were larger than 1 mm in diameter. Transcriptomic analysis revealed that the greatest gene expression changes occurred when GBOs were 2 mm in diameter, with collagen VI as the most upregulated ECM-related gene. Quantitative and histochemical assessments further supported native ECM synthesis with significantly higher levels of glycosaminoglycans and collagen in GBOs compared with spheroids. To our knowledge, this study presents the first reproducibly large GBOs with natively produced ECMs. Organoids with natively synthesized ECMs promise to eliminate artifacts and variability from aged, homogeneic, or xenogeneic scaffolds and to provide insights for ECM-targeted drug development.

Overview

  • bulletstudy, to generate glioblastoma multiforme (GBM) organoids (GBOs) without exogenous extracellular matrices (ECMs) and without cell culture media changes.
  • bulletmethod: stem cell differentiation without encapsulation in Matrigel™ or other ECMs, and without media changes to produce stable tissue-like GBOs.
  • bulletpurpose: to develop a reproducible method for generating large GBOs with natively produced ECMs, eliminating artifacts and variability from aged, homogeneous, or xenogeneic scaffolds.

Comparative Analysis & Findings

  • bulletthe study observed a transition from homogenous cell populations to tissue-like structures when GBOs were larger than 1 mm in diameter.
  • bullethes greatest gene expression changes occurred when GBOs were 2 mm in diameter, with collagen VI as the most upregulated ECM-related gene.
  • bulletquantitative and histochemical assessments supported native ECM synthesis with significantly higher levels of glycosaminoglycans and collagen in GBOs compared with spheroids.

Implications and Future Directions

  • bulletthis study presents a reproducibly large GBOs with natively produced ECMs, promising to eliminate artifacts and variability from aged, homogeneous, or xenogeneic scaffolds.
  • bulletthese GBOs could provide insights for ECM-targeted drug development.
  • bulletrand could be a valuable tool for understanding the dynamic reciprocity between cells and ECMs in tissue development and the role of ECMs in cancer progression.