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Improving expansion of pluripotent human embryonic stem cells in perfused bioreactors through oxygen control.

Abstract：

:The successful transfer of human embryonic stem cell (hESC) technology and cellular products into clinical and industrial applications needs to address issues of automation, standardization and the generation of relevant cell numbers of high quality. In this study, we combined microcarrier technology and controlled stirred tank bioreactors, to develop an efficient and scalable system for expansion of pluripotent hESCs. We demonstrate the importance of controlling pO(2) at 30% air saturation to improve hESCs growth. This concentration allowed for a higher energetic cell metabolism, increased growth rate and maximum cell concentration in contrast to 5% pO(2) where a shift to anaerobic metabolism was observed, decreasing cell expansion 3-fold. Importantly, the incorporation of an automated perfusion system in the bioreactor enhanced culture performance and allowed the continuous addition of small molecules assuring higher cell concentrations for a longer time period. The expanded hESCs retained their undifferentiated phenotype and pluripotency. Our results show, for the first time, that the use of controlled bioreactors is critical to ensure the production of high quality hESCs. When compared to the standard colony culture, our strategy improves the final yield of hESCs by 12-fold, providing a potential bioprocess to be transferred to clinical and industrial applications.

journal_name

J Biotechnol

journal_title

Journal of biotechnology

authors

Serra M,Brito C,Sousa MF,Jensen J,Tostões R,Clemente J,Strehl R,Hyllner J,Carrondo MJ,Alves PM

doi

10.1016/j.jbiotec.2010.06.015

subject

Has Abstract

pub_date

2010-08-02 00:00:00

pages

208-15

issue

4

eissn

0168-1656

issn

1873-4863

pii

S0168-1656(10)00275-0

journal_volume

148

pub_type

杂志文章

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