Shaken bioreactors in the field of cell cultivation

Anderlei T, Kühner M, Kühner AG, SwitzerlandBüchs J, BioVT, RWTH Aachen, GermanyStettler M, de Jesus M, Wurm F, EPFL, Switzerland

Adolf Kühner AG, Dinkelbergstr. 1, CH-4127 Birsfelden (Basel),Switzerland, fon: +41 61 319 9393, www.kuhner.com

orbitalshaking

20mL

100mL

10L

100L

1000L

1mL

Good Scalability:

180 RPM50 mm

200 RPM50 mm

220 RPM50 mm

+ same hydrodynamic+ surface aeration+ calculable+ no foam+ no bubbles

Orbital shaking technology has been widely used in the biotechnological field because of easy-of-use, increased flexibility and reduced costs. Recently, orbital shaken bioreactors of scales from mL - 1000L have been employed for cultivation of mammalian cells and are expected to become attractive alternatives to conventional stirred-tank bioreactors. To meet the need of developing a mature bioreactor system, extensive investigations, including simulations using Computational Fluid Dynamics (CFD), are underway to better understand the special engineering properties resulted from orbital shaking.

20mL 12L

1.00e-01

8.01e-02

6.01e-02

4.02e-02

2.02e-02

2.89e-04

Pa

30-L cylinder, working volume 12L,shaking speed 80 rpmShear stress is two logs

below cell damage

4.35e+00

8.71e-01

1.74e+00

2.61e+00

3.48e+00

1.15e-04

Pa

parameter small scale large scale optimisation shear stress low low none

evaporation high low humidified environment

aeration

high

middle

oxygen enriched air helical track

Evaporation: 250mL narrow necked shake flask

oxygen transfer rate in microtiterplates

Comparison: 50mL tube and 55L cylindrical vessel with helical track

0

0.01

0.02

0.03

0.04

0.05

0.06

30 40 50 60 70 80 90 100

relative humidity [%]

ev

ap

ora

tio

n[g

/h]

37°C (Corning)

37°C

30°C

25°C

18°C

0,00

0,02

0,04

0,06

0,08

0,10

0,12

0,14

0,16

0,18

0,20

0 200 400 600 800 1000

shaking frequency [rpm]

Deep-Well squareDeep-Well roundMTP Standard round

d0= 25 mm, VL= 200µl

Cell density_HTCell density_50mL tubeViability_50mL tubeViability_HT

Time [h]0

050

202

4

6

8

10

12

40

60

80

100

100

1500

bt

[]

Via

iliy

%

6b

td

sit

[0

cll

/]

Via

iliy

en

y 1

e

sm

L