A Comparative Study of Suspension Cultivation Systems for the Expansion of Undifferentiated Mouse Embryonic Stem Cells

Abstract

Embryonic Stem (ES) cells are self- renewing and pluripotent cells that can differentiate into a variety of cell lineages. Implementation in tissue engineering or as model systems for drug discovery makes ES cells attractive as a cell source. To use ES cells for these applications, technologies are required to generate a large number of cells with defined characteristics. Currently, adherent culture methods are routinely applied for the maintenance of undifferentiated ES cells. However, perfused and stirred bioreactors enable a more homogenous environment and, more importantly, facilitate the ability to monitor and control culture parameters (e.g. oxygen content and pH) which is advantageous compared with static culture vessels. In this work, four different suspension cultivation systems (two spinner flasks with different stirrer design, an Erlenmeyer flask and a Petri dish) were compared with respect to the ability to generate undifferentiated ES cells. Therefore, the murine ES cell line E14.1, 129/Ola with an eGFP transgene targeted to the Brachyury locus was used. Besides the evaluation of biomass production, pluripotency marker expression was analyzed applying flow cytometry.