3. Freshly sterilized conditioned Cytodex 3 microcarriers (Cytiva,

USA).

4. Resazurin-based PrestoBlue™assay (PrestoBlue, Thermo

Fisher Scientific, USA).

2.4.4

Glucose

Consumption Rate

1. UV-VIS spectrophotometer GENESYS 20 (Thermo Fisher

Scientific, USA).

2. BioMaxima-glucose enzymatic assay (BioMaxima, Poland).

3. Double-distilled water.

2.4.5

Activity of Lactate

Dehydrogenase (LDH)

1. UV-VIS spectrophotometer GENESYS 20 (Thermo Fisher

Scientific, USA).

2. BioMaxima-LDH enzymatic assay (BioMaxima, Poland).

3. Double-distilled water.

2.5

General

Description of the

Setup

2.5.1

ReadyToProcess

WAVE^™25 Bioreactor

System

The schematic diagram of ReadyToProcess WAVE^TM25 system

(WAVE 25, Cytiva, USA) setup has been presented in Fig. 1.

WAVE 25 was equipped with a disposable pre-sterilized polymer-

based 2 L Cellbag™bag-like container (Cellbag, Cytiva, USA),

which is characterized by working volume ranging from 0.1 L to

1.0 L of culture medium, as recommended by supplier of the

system. The schematic diagram of Cellbag container and its ports

configuration has been presented in Fig. 2. The thermostatic rocker

performed continuously oscillating movements of whole Cellbag

container, according to variously set conditions of wave-type agita-

tion defined by: angle of oscillation, frequency of oscillation, and

rocking motion acceleration. The control unit (CBCU) allowed to

automatic measuring and controlling of crucial parameters of cul-

ture

conditions,

such

as

pH

level

and

dissolved

oxygen

(DO) concentration (via miniaturized spectrophotometric sensors

Fig. 1 Schematic diagram of WAVE 25 bioreactor system

Microcarrier-Supported Culture of Chondrocytes in Disposable Bioreactor

149