Microcarrier-Supported Culture of Chondrocytes in Continuously Rocked Disposable Bioreactor

Methods Mol Biol. 2022:2436:145-156. doi: 10.1007/7651_2021_411.

Abstract

Disposable wave-assisted bioreactors are devices originally designed for scaling-up cultures of extremely fragile animal cells. In such bioreactors, agitation is achieved by continuous horizontal oscillations of disposable culture bag-like container fixed in a rocker unit. The continuous rocking movement of the container induces waves in the two-phase (i.e., gas-liquid) culture system composed of CO2-enriched air and aqueous culture medium. Such continuously oscillating devices can be utilized for supporting homogeneity in systems for in vitro propagation of animal anchorage-dependent, that is, adherent, cells, like CP5 chondrocytes cells. As most of in vitro cultured cells exhibit anchorage-dependency toward solid surface, the suitable interface can be provided by beads of microcarriers made of polymers and characterized by large surface-to-volume ratio. This chapter describes a methodology for efficient propagation of CP5 chondrocytes on Cytodex 3 microcarriers performed in ReadyToProcess WAVE 25 disposable bioreactor, as well as all useful procedures for daily monitoring the growth of CP5 chondrocytes.

Keywords: Anchorage-dependent cells; Bioprocess intensification; Chondrocytes; Disposable bioreactor; Microcarrier beads; Wave-assisted agitation.

MeSH terms

  • Animals
  • Bioreactors
  • Cell Culture Techniques* / methods
  • Cell Line
  • Cells, Cultured
  • Chondrocytes*