Protocol for in vitro evaluation of effects of stiffness on patient-derived glioblastoma

STAR Protoc. 2024 Sep 20;5(3):103266. doi: 10.1016/j.xpro.2024.103266. Epub 2024 Aug 14.

Abstract

Glioblastoma (GBM) is the most common and lethal type of primary brain tumor. Physiologically, GBM cells experience a heterogeneous mechanical landscape. Here, we present an in vitro method to study the effects of tissue stiffness on patient-derived GBM that utilizes hyaluronic acid (HA)-based, mechanically tunable scaffolds for three-dimensional (3D) culture of patient-derived GBM spheroids. We describe steps to fabricate and characterize HA-based scaffolds, culture GBM spheroids within 3D hydrogel scaffolds, and prepare cultured cells for a variety of experimental assessments. For complete details on the use and execution of this protocol, please refer to Sohrabi et al.1.

Keywords: Cancer; Material sciences; Tissue Engineering.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Culture Media* / chemical synthesis
  • Glioblastoma*
  • Humans
  • Hyaluronic Acid
  • Hydrogels / chemical synthesis
  • Primary Cell Culture* / methods
  • Spheroids, Cellular

Substances

  • Hydrogels
  • Hyaluronic Acid
  • Culture Media