for large-scale human embryonic stem cell pro-

duction. Cell Death Dis 9. https://doi.org/

10.1038/s41419-018-0863-8

6. Nogueira DES, Rodrigues CAV, Carvalho MS,

Miranda CC, Hashimura Y, Jung S, Lee B,

Cabral JMS (2019) Strategies for the expan-

sion of human induced pluripotent stem cells as

aggregates in single-use Vertical-Wheelbior-

eactors. J Biol Eng 13. https://doi.org/10.

1186/s13036-019-0204-1

7. Shafa

M,

Panchalingam

KM,

Walsh

T,

Richardson

T,

Baghbaderani

BA

(2019)

Computational fluid dynamics modeling, a

novel, and effective approach for developing

scalable cell therapy manufacturing processes.

Biotechnol Bioeng 116:3228–3241

8. Pan T, Chen Y, Zhuang Y, Yang F, Xu Y, Tao J,

You K, Wang N, Wu Y, Lin X, Wu F, Liu Y,

Li Y, Wang G, Li Y, Wang G, Li Y-X (2019)

Synergistic modulation of signaling pathways

to expand and maintain the bipotency of

human hepatoblasts. Stem Cell Res Ther 10.

https://doi.org/10.1186/s13287-019-1463-

y

9. Pandey PR, Tomney A, Woon MT, Uth N,

Shafighi

F,

Ngabo

I,

Vallabhaneni

H,

Levinson Y, Abraham E, Ben-Nun IF (2020)

End-to-end platform for human pluripotent

stem cell manufacturing. Int J Mol Sci 21.

https://doi.org/10.3390/ijms21010089

10. Jensen C, Teng Y (2020) Is it time to start

transitioning from 2D to 3D cell culture?

Front

Mol

Biosci

7.

https://doi.org/10.

3389/fmolb.2020.00033

11. Edmondson R, Broglie JJ, Adcock AF, Yang L

(2014) Three-dimensional cell culture systems

and their applications in drug discovery and

cell-based biosensors. Assay Drug Dev Technol

12:207–218

12. Langhans

SA

(2018)

Three-dimensional

in vitro cell culture models in drug discovery

and

drug

repositioning.

Front

Pharmacol

9.

https://doi.org/10.3389/fphar.2018.

00006

13. Lv D, Hu Z, Lu L, Lu H, Xu X (2017) Three-

dimensional cell culture: a powerful tool in

tumor research and drug discovery. Oncol

Lett 14:6999–7010

14. Mummery CL, Zhang J, Ng ES, Elliott DA,

Elefanty AG, Kamp TJ (2012) Differentiation

of human embryonic stem cells and induced

pluripotent stem cells to cardiomyocytes: a

methods overview. Circ Res 111:344–358

15. Schwedhelm I, Zdzieblo D, Appelt-Menzel A,

Berger C, Schmitz T, Schuldt B, Franke A,

Mu¨ller FJ, Pless O, Schwarz T, Wiedemann P,

Walles H, Hansmann J (2019) Automated real-

time monitoring of human pluripotent stem

cell aggregation in stirred tank reactors. Sci

Rep

9.

https://doi.org/10.1038/s41598-

019-48814-w

16. Badenes SM, Fernandes TG, Cordeiro CSM,

Boucher S, Kuninger D, Vemuri MC, Diogo

MM, Cabral JMS (2016) Defined essential

80 medium and vitronectin efficiently support

scalable xeno-free expansion of human induced

pluripotent stem cells in stirred microcarrier

culture systems. PLoS One 11. https://doi.

org/10.1371/journal.pone.0151264

Expansion of Human Pluripotent Stem Cells in Stirred Tank Bioreactors

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