Improved Oxygen Supply to Multicellular Spheroids Using A Gas-permeable Plate and Embedded Hydrogel Beads

Cells. 2019 May 31;8(6):525. doi: 10.3390/cells8060525.

Abstract

Culture systems for three-dimensional tissues, such as multicellular spheroids, are indispensable for high-throughput screening of primary or patient-derived xenograft (PDX)-expanded cancer tissues. Oxygen supply to the center of such spheroids is particularly critical for maintaining cellular functions as well as avoiding the development of a necrotic core. In this study, we evaluated two methods to enhance oxygen supply: (1) using a culture plate with a gas-permeable polydimethylsiloxane (PDMS) membrane on the bottom, and; (2) embedding hydrogel beads in the spheroids. Culturing spheroids on PDMS increased cell growth and affected glucose/lactate metabolism and CYP3A4 mRNA expression and subsequent enzyme activity. The spheroids, comprised of 5000 Hep G2 cells and 5000 20 µm-diameter hydrogel beads, did not develop a necrotic core for nine days when cultured on a gas-permeable sheet. In contrast, central necrosis in spheroids lacking hydrogel beads was observed after day 3 of culture, even when using PDMS. These results indicate that the combination of gas-permeable culture equipment and embedded hydrogel beads improves culture 3D spheroids produced from primary or PDX-expanded tumor cells.

Keywords: 3D culture; gas-permeable plate; hydrogel beads; methylcellulose; multicellular spheroids.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus / drug effects
  • Cell Nucleus / metabolism
  • Cell Proliferation / drug effects
  • Cytochrome P-450 CYP3A / genetics
  • Cytochrome P-450 CYP3A / metabolism
  • DNA / metabolism
  • Energy Metabolism / drug effects
  • Gases / chemistry*
  • Gene Expression Regulation, Enzymologic / drug effects
  • Glucose / metabolism
  • Hep G2 Cells
  • Humans
  • Hydrogels / pharmacology*
  • Ki-67 Antigen / metabolism
  • Microspheres*
  • Necrosis
  • Nitroimidazoles / pharmacology
  • Oxygen / pharmacology*
  • Permeability
  • Spheroids, Cellular / drug effects*

Substances

  • Gases
  • Hydrogels
  • Ki-67 Antigen
  • Nitroimidazoles
  • pimonidazole
  • DNA
  • Cytochrome P-450 CYP3A
  • Glucose
  • Oxygen