Large-scale culture of a megakaryocytic progenitor cell line with a single-use bioreactor system

Retno Wahyu Nurhayati, Yoshihiro Ojima, Takeaki Dohda, Masahiro Kino-Oka

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

The increasing application of regenerative medicine has generated a growing demand for stem cells and their derivatives. Single-use bioreactors offer an attractive platform for stem cell expansion owing to their scalability for large-scale production and feasibility of meeting clinical-grade standards. The current work evaluated the capacity of a single-use bioreactor system (1 L working volume) for expanding Meg01 cells, a megakaryocytic (MK) progenitor cell line. Oxygen supply was provided by surface aeration to minimize foaming and orbital shaking was used to promote oxygen transfer. Oxygen transfer rates (kLa) of shaking speeds 50, 100, and 125 rpm were estimated to be 0.39, 1.12, and 10.45 h−1, respectively. Shaking speed was a critical factor for optimizing cell growth. At 50 rpm, Meg01 cells exhibited restricted growth due to insufficient mixing. A negative effect occurred when the shaking speed was increased to 125 rpm, likely caused by high hydrodynamic shear stress. The bioreactor culture achieved the highest growth profile when shaken at 100 rpm, achieving a total expansion rate up to 5.7-fold with a total cell number of 1.2 ± 0.2 × 109 cells L−1. In addition, cells expanded using the bioreactor system could maintain their potency to differentiate following the MK lineage, as analyzed from specific surface protein and morphological similarity with the cells grown in the conventional culturing system. Our study reports the impact of operational variables such as shaking speed for growth profile and MK differentiation potential of a progenitor cell line in a single-use bioreactor.

Original languageEnglish
Pages (from-to)362-369
Number of pages8
JournalBiotechnology Progress
Volume34
Issue number2
DOIs
Publication statusPublished - 1 Mar 2018

Keywords

  • bioreactor
  • cell differentiation
  • large-scale
  • megakaryocyte
  • single-use bag

Fingerprint

Dive into the research topics of 'Large-scale culture of a megakaryocytic progenitor cell line with a single-use bioreactor system'. Together they form a unique fingerprint.

Cite this