2.1

Microcarrier

Conditioning

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

Scientific, USA).

2. Laboratory autoclave (Biolab Scientific Ltd., Canada).

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

China).

4. Manual pipette controller and presterilized disposable serolog-

ical pipettes (25 mL, 10 mL, and 5 mL).

5. Automatic laboratory pipettes and presterilized disposable

pipette tips (1 mL and 200 μL) placed in the racks.

6. 50 mL Falcon centrifuge tubes.

7. Cytodex 3 (Cytiva, USA), as microcarriers made of pig gela-

tine–coated cross-linked dextran beads, characterized by diam-

eter equaled to 175 μm, surface area dry weight equaled to

0.27 m² g
¹ and beads quantity in dry weight at level of

4.3  10⁶ g
¹.

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

cations (DPBS, Thermo Fisher Scientific, USA).

9. 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).

2.2

Preparation of

CP5 Cell Inoculum

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

Scientific, USA).

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

China).

3. Laboratory centrifuge for Falcon tubes MPW-352RH (MPW,

Poland).

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

5. Disposable Cellbag™2L vessel compatible with ReadyToProcess

WAVE™25 system (Cellbag, Cytiva, USA).

6. Manual pipette controller and pre-sterilized disposable sero-

logical pipettes (10 mL and 5 mL).

7. Automatic laboratory pipettes and presterilized pipette tips

(1 mL and 200 μL) placed in the racks.

8. 50 mL Falcon centrifuge tubes.

9. Articular cartilage progenitor cell line isolated from Bos taurus

(CP5, ECACC, UK).

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

cations (DPBS, Thermo Fisher Scientific, USA).

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

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

Microcarrier-Supported Culture of Chondrocytes in Disposable Bioreactor

147