inoculation................................................................... 9

metabolite analyzers.................................................. 11

paper autoclaving bags.............................................. 12

parameters ................................................................... 6

probe calibration and sterilization.............................. 7

probes .......................................................................... 5

sampling port .............................................................. 4

seeding cell density.................................................... 13

supplements................................................................. 6

tubing welders........................................................... 11

Mature cardiomyocytes (CM)........................................ 67

Mechanical stimulation.................................................128

Mechanobiology ...........................................................135

Media composition ......................................................... 85

Medium adaptability ....................................................... 88

Membrane-based technologies.....................................257

Microbial continuous cultivation .................................224

Microcarrier-based cell culture.....................................257

Microcarrier beads ...................................... 146, 151, 155

Microcarrier spinner cultures

cardiac differentiation in.....................................75–77

media and small molecules for ................................. 69

preparation of......................................................70–71

proper suspension of................................................. 77

reagents and consumables for .................................. 69

in suspension and ensure adequate mass

transfer ........................................................... 85

MTT assay .....................................................................245

Mycoplasma............................................................ 88, 191

P

Partial decellularization.................................................168

Patellar tendon..............................................................138

PBS vertical wheel (PBS-VW) bioreactors..................194,

196, 199, 200

PEG6000..................................................... 196, 201, 202

Perfusion............................................................... 242, 243

Poisson distribution ........................................................ 85

Polydimethylsiloxane (PDMS)............................ 260–265

Polyethylene terephthalate (PET)................................190

Polytetrafluoroethylene (PTFE)-coated magnetic stir

bar.................................................................243

Population doubling level (PDL) .................................. 90

PureLink^™RNA Mini Kit ............................................143

R

ReadyToProcess WAVE 25 disposable

bioreactor............................................ 146–150

Recellularization............................................................167

Rocked disposable bioreactor, chondrocytes

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 WAVE^TM 25 bioreactor

system.................................................. 149–150

rehydration, conditioning, and sterilization of

microcarriers ................................................151

samples for analysis ........................................ 148, 152

Rotating bioreactor..................................... 158, 159, 161

RT-qPCR analysis .........................................................142

S

Saccharomyces cerevisiae cells........................................... 37

Scaffold-based method .................................................136

Shed cells ..................................................... 187, 188, 191

Single-use bioreactors..................................................... 84

Sodium dodecyl sulfate (SDS)............................. 168, 172

Spheroid-based cultures.................................................. 87

Stem cell expansions ....................................................... 90

Stirred bioreactor system ..............................................243

Stirred tank reactors (STR)

DO concentration

aeration ................................................................ 18

bioreactor.......................................................19, 20

chronometer........................................................ 20

dynamic method ...........................................23, 24

electrode of.......................................................... 24

growth media ...................................................... 20

microorganism ....................................... 19, 21, 23

oxygen transfer coefficient............................21–22

spectrophotometer.............................................. 20

steady-state technique......................................... 19

sterilization process............................................. 21

unsteady-state technique ..............................18–19

human pluripotent stem cells

autoclave cycle..................................................... 50

bioreactor.......................................................41–42

bioreactor setup.............................................44–46

buffers and reagents............................................ 40

cardiomyoyctes.................................................... 40

cell-based therapies ............................................. 39

cell count during expansion .........................47–48

cell harvest .....................................................48–49

cell inoculum ....................................................... 46

BIOREACTORS IN STEM CELL BIOLOGY: METHODS AND PROTOCOLS

Index ²⁷¹