of inactivated fetal bovine serum (FBS, Thermo Fisher Scien-

tific, USA), and 1% of commercial antibiotic–antimycotic mix-

ture (PenStrep, Thermo Fisher Scientific, USA).

12. 0.05% trypsin–EDTA containing phenol-red (Thermo Fisher

Scientific, USA).

2.3

Cell Culture

1. ReadyToProcess

WAVE™

25

(WAVE

25,

Cytiva,

USA)

bioreactor system.

2. Disposable Cellbag™2L bag-like container adapted for WAVE

25 system.

3. Disposable 5 mL Luer Lock tip syringe (BD, Germany).

2.4

Analytical

Methods

2.4.1

Preparation of

Samples for Analysis

1. Biological safety cabinet Herasafe^™KS 12 (Thermo Fisher

Scientific, USA).

2. Mixer Vortex MX-S (Biologix Group, People’s Republic of

China).

3. Pipette controller, disposable serological pipettes (25 mL,

10 mL, and 5 mL), laboratory pipettes, pipette tips (1 mL

and 200 μL), and pipette aids.

4. 2 mL Eppendorf tubes.

5. Syringe filter φ ¼ 0.2 μm WhatMan^® Puradisc (Thermo Fisher

Scientific, USA).

6. Dulbecco’s phosphate-buffered saline free of Ca²⁺ and Mg²⁺

cations (DPBS, Thermo Fisher Scientific, USA).

7. Dulbecco’s Modified Eagle’s Medium (DMEM, Thermo

Fisher Scientific, USA) containing 4.5 g L
¹ of glucose, 10%

of inactivated fetal bovine serum (FBS, Thermo Fisher Scien-

tific, USA), and 1% of commercial antibiotic–antimycotic mix-

ture (PenStrep, Thermo Fisher Scientific, USA).

8. 0.05% trypsin–EDTA with phenol red (Thermo Fisher Scien-

tific, USA).

2.4.2

Cell Staining

1. Reverse microscope Eclipse TS100 (Nikon, Japan).

2. Bu¨rker-Tu¨rk haemocytometer (Brand, Germany).

3. 0.4% trypan blue aqueous solution (Thermo Fisher Scientific,

USA).

2.4.3

Activity of

Intracellular

Oxidoreductases

1. CO2 incubator HF90 (Heal Force, People’s Republic of

China).

2. UV-VIS spectrophotometer GENESYS 20 (Thermo Fisher

Scientific, USA).

148

Kamil Wierzchowski and Maciej Pilarek