preculture and batch phase.......................229–230
product determinatio........................................ 235
substrate uptake rates........................................ 235
Cedex Bio Analyzer.......................................................105
CelGro® scaffold .................................................. 136, 140
Cell activity, in 3D scaffolds
bioartificial tissues ...................................................127
bioreactor system ........................................... 129–130
mechanical compression load and fluid flow
parameters....................................................131
sample preparation ..................................................130
sample transfer and chamber assembly ..................131
Cell adhesion................................................................... 85
Cell attachment and matrix synthesis ..........................131
Cell attachment efficiency............................................... 88
Cellbag container ................................149–152, 154, 155
Cell colonization studies, bacterial nanocellulose-based
grafts for.............................................. 205–207
materials
antibodies .......................................................... 208
assays and staining solutions............................. 208
bioreactor setup.........................................207–208
cell culture ......................................................... 208
chemicals and solutions .................................... 208
consumables ...................................................... 209
hardware ....................................................209–210
methods
AcLDL uptake assay.......................................... 216
acridine orange staining.................................... 218
bioreactor setup.........................................210–211
CD31 immunofluorescence staining .......218–219
cell seeding ................................................211–212
considerations.................................................... 210
lactate monitoring.....................................212–213
Phalloidin—FActin staining .....................216–218
small diameter vascular grafts, connection ...... 211
termination of experiment................................ 216
WST-1 proliferation assay.........................213–215
Cell culture medium composition ................................. 88
Cell expansion ...........................................................73–75
Cell growth...................................................................... 85
Cell harvesting...............................................................260
CELLine AD 1000 bioreactor flask
adapting cells, to long-term bioreactor
media................................................... 186–187
cell culture ...............................................................185
characterization materials .......................................185
EVs
characterization .........................................188–189
harvesting .......................................................... 187
isolation and purification ......................... 188, 191
isolation materials.............................................. 185
RNA................................................................... 190
illustration................................................................184
imaging bioreactor growth surface ............... 189–190
inoculation of cells ......................................... 185–186
preparation ..............................................................185
shed cells......................................................... 187, 188
ultrafiltration or multiple ultracentrifuge
spins..............................................................183
Centrifugation...................................................... 197, 199
Chemostat cultivation.......................................... 223, 224
Chinese hamster ovary (CHO) cells ................................ 1
Chondrocytes, rocked disposable bioreactor
advantages................................................................145
cellbag container with inlet and outlet port..........150
cell culture ...............................................................148
cell density and viability..........................................153
cell staining..............................................................148
CP5 cell inoculum preparation ..................... 147–148
DO and pH level.....................................................154
glucose consumption rate......................149, 153–154
intracellular oxidoreductases ......................... 148–149
lactate dehydrogenase.................................... 149, 154
maintaining CP5 cells in bioreactor.......................152
metabolic activity of cells adhered to
microcarriers ................................................153
microcarrier conditioning.......................................147
preparation of CP5 chondrocyte
inoculum............................................. 151–152
ReadyToProcess WAVETM 25 bioreactor
system.................................................. 149–150
rehydration, conditioning, and sterilization of
microcarriers ................................................151
samples for analysis ........................................ 148, 152
Classic fed-batch cultivation............................................. 2
CLAVE™sampling port ...............................................155
Commercially available single-use bioreactors .............. 84
Compression bioreactors ..............................................128
Conventional continuous process times ......................224
CP5 chondrocytes...............................................146, 147,
151, 152, 154, 155
Cryovials ............................................................................ 2
Cultivation container ...................................................... 84
Cytodex 3 microcarriers .....................146, 147, 149, 151
D
Decellularization .................................158, 164, 167, 168
Decellularized tracheal ECM-based porcine bio-scaffold
generation....................................................... 160–163
repopulation ...........................................161–162, 164
Detergent circuitries ............................................ 173–174
Dissolved oxygen (DO) concentration, STR
aeration ...................................................................... 18
bioreactor.............................................................19, 20
chronometer.............................................................. 20
dynamic method .................................................23, 24
electrode of................................................................ 24
268 BIOREACTORS IN STEM CELL BIOLOGY: METHODS AND PROTOCOLS
Index