One of the key challenge
s in
stem cell bioproce
ssing i
s the large-
scale cultivation of
stem cell
s in order to meet the demanding meaningful cell number
s needed for biomedical application
s, e
specially for clinical
setting
s. Mou
se embryonic
stem cell
s , u
sed a
s a model
sy
stem herein, were cultivated on microcarrier
s in a fully controlled
stirred tank reactor (STR) . The impact of varying the concentration of di
ssolved oxygen (at 5%, 10%, 20% and 30% DO) and operating under a continuou
s perfu
sion mode on cell growth and pluripotency maintenance wa
s inve
stigated. In addition, in order to further optimize the feeding
strategy of the STR operating under continuou
s perfu
sion toward maximal cell production, the influence of different medium re
sidence
s time
s (12 h, 24 h, 32 h, 48 h and 96 h) wa
s evaluated. Overall, the maximal cell concentration of 7.9-9.2 脳 10<
sup>6
sup> cell
s/mL were attained after 11 day
s, with no pa
ssaging required, under a DO of 10-20% in the continuou
s perfu
sed bioreactor with cell retention and medium re
sidence
s time
s of 32-48 h. Importantly, mESC expanded under the
se condition
s, retained the expre
ssion of pluripotency marker
s (Oct4, Nanog and S
sea-1), a
s well a
s their differentiation potential into cell
s of the three embryonic germ layer
s.
The STR-based cultivation platform optimized herein represents a major contribution toward the development of large-volume production systems of differentiated cell derivatives for a wide range of biomedical applications.