12. 15-mL and/or 50-mL centrifuge tube (Sarstedt, Nu ¨mbrecht,
Germany).
13. Aspiration system (Vacusafe, Integra, Biebertal, Germany).
14. Centrifuge (Heraeus Megafuge X1R, Thermo Fisher Scientific,
Darmstadt, Germany).
15. Incubator at 37
C (HERAcell
® 240i CO 2 Incubator, Thermo
Fisher Scientific, Waltham, USA).
16. Incident light microscope (DM1i, Leica, Wetzlar, Germany).
2.1.3 Cell Counting
and Cell Viability
Determination via
a Counting Chamber
1. Hemocytometer/counting chamber, Neubauer improved
(Marienfeld, Ko ¨nigshofen, Germany).
2. PBS (#L1825, Biochrom, Berlin, Germany).
3. Incident light microscope (DM1i, Leica, Wetzlar, Germany).
4. Pipette and pipette tips (Sarstedt, Nu ¨mbrecht, Germany).
5. 0.4% trypan blue solution (Sigma-Aldrich, Hamburg,
Germany).
2.1.4 Cell Counting
and Cell Viability
Determination via Crystal
Violet
1. Centrifuge (Heraeus Megafuge X1R, Thermo Fisher Scientific,
Darmstadt, Germany).
2. 1.5 mL tubes (Eppendorf, Hamburg, Germany).
3. 0.1% (w/v) crystal violet staining solution (Carl Roth, Karlsruhe, Germany).
4. 0.1 M citric acid (Carl Roth, Karlsruhe, Germany).
5. Hemocytometer/counting chamber, Neubauer improved
(Marienfeld, Ko ¨nigshofen, Germany).
6. Incident light microscope (DM1i, Leica, Wetzlar, Germany).
2.1.5 Bioreactor
Preparation
1. 1-L glass STR, working volume: 0.5 L (Z611000110; Applikon, Biotechnology, Delft, Netherlands).
2. Software: Process control (BioPAT
® MFCS SCADA, Sartorius
AG, Go ¨ttingen, Germany).
3. Reactor control unit (BIOSTAT
® B-DCU Twin, Sartorius
Stedim Biotech GmbH, Go ¨ttingen, Germany).
4. 3 Â 45
pitched-blade impeller, diameter: 4.5 cm (Sartorius
Stedim Biotech GmbH, Go ¨ttingen, Germany).
5. Sparger for aeration (Sartorius Stedim Biotech GmbH, Go ¨ttingen, Germany).
6. Sensors for monitoring:
(a) pH sensor (Z001023551, Applikon Biotechnology, JG
Delft, Netherlands).
(b) PT100 temperature sensor (Sartorius Stedim Biotech
GmbH, Go ¨ttingen, Germany).
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Germany).
13. Aspiration system (Vacusafe, Integra, Biebertal, Germany).
14. Centrifuge (Heraeus Megafuge X1R, Thermo Fisher Scientific,
Darmstadt, Germany).
15. Incubator at 37
C (HERAcell
® 240i CO 2 Incubator, Thermo
Fisher Scientific, Waltham, USA).
16. Incident light microscope (DM1i, Leica, Wetzlar, Germany).
2.1.3 Cell Counting
and Cell Viability
Determination via
a Counting Chamber
1. Hemocytometer/counting chamber, Neubauer improved
(Marienfeld, Ko ¨nigshofen, Germany).
2. PBS (#L1825, Biochrom, Berlin, Germany).
3. Incident light microscope (DM1i, Leica, Wetzlar, Germany).
4. Pipette and pipette tips (Sarstedt, Nu ¨mbrecht, Germany).
5. 0.4% trypan blue solution (Sigma-Aldrich, Hamburg,
Germany).
2.1.4 Cell Counting
and Cell Viability
Determination via Crystal
Violet
1. Centrifuge (Heraeus Megafuge X1R, Thermo Fisher Scientific,
Darmstadt, Germany).
2. 1.5 mL tubes (Eppendorf, Hamburg, Germany).
3. 0.1% (w/v) crystal violet staining solution (Carl Roth, Karlsruhe, Germany).
4. 0.1 M citric acid (Carl Roth, Karlsruhe, Germany).
5. Hemocytometer/counting chamber, Neubauer improved
(Marienfeld, Ko ¨nigshofen, Germany).
6. Incident light microscope (DM1i, Leica, Wetzlar, Germany).
2.1.5 Bioreactor
Preparation
1. 1-L glass STR, working volume: 0.5 L (Z611000110; Applikon, Biotechnology, Delft, Netherlands).
2. Software: Process control (BioPAT
® MFCS SCADA, Sartorius
AG, Go ¨ttingen, Germany).
3. Reactor control unit (BIOSTAT
® B-DCU Twin, Sartorius
Stedim Biotech GmbH, Go ¨ttingen, Germany).
4. 3 Â 45
pitched-blade impeller, diameter: 4.5 cm (Sartorius
Stedim Biotech GmbH, Go ¨ttingen, Germany).
5. Sparger for aeration (Sartorius Stedim Biotech GmbH, Go ¨ttingen, Germany).
6. Sensors for monitoring:
(a) pH sensor (Z001023551, Applikon Biotechnology, JG
Delft, Netherlands).
(b) PT100 temperature sensor (Sartorius Stedim Biotech
GmbH, Go ¨ttingen, Germany).
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