Human mesenchymal stromal/stem cells

(hMSCs) (cont.)

expansion in planar and bioreactor

cultures................................................ 195–196

expansion in planar culture.....................................199

Human pluripotent stem cells (hPSCs)

random positioning machine

APLNR⁺ mesoderm cells.................................... 56

bioreactor equipment ......................................... 57

cell culture medium and reagents ...................... 57

cell lines ............................................................... 57

electronic control system.................................... 65

feeder-free expansion .......................................... 59

flow cytometry .................................................... 64

hematopoietic differentiation.......................55, 56

hematopoietic progenitor cells.....................61–62

hemogenic endothelium progenitor............60–61

IF9S medium formulation.................................. 58

immunofluorescence staining.......................62–64

immunostaining and flow cytometry................. 59

Matrigel^® solution .............................................. 64

mesoderm induction........................................... 60

T12.5 cell culture flasks ...................................... 59

stirred tank bioreactors

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

C-flex tubing ....................................................... 46

DeltaV control platform ..................................... 51

DO probe ............................................................ 51

extracellular matrix coating ................................ 43

F3 hPSC passaging solution.........................49, 52

harvest line extension.......................................... 42

L7^™culture system................................ 40, 49, 50

microcarrier coating............................................ 44

passaging solution............................................... 40

Pharmed tubing portion..................................... 47

T-75 flask............................................................. 43

tubing ............................................................40–41

2D seed train .................................................43–44

using peristaltic pump......................................... 52

Human umbilical cord-derived MSCs

(hUC-MSCs) ...................................... 194, 200

Human umbilical cord MSC-secreted EVs .................194

Human umbilical vein endothelial cells

(HUVECs)...................................................194

I

Inertial microfluidics.....................................................258

Intestinal mucosa ..........................................................246

Intestinal tissue model ..................................................248

In vitro bioreactor perfusion model ................... 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

In vitro 3D mechanical stimulation

of TDSCs ............................................ 135–141

Isopropyl-α-D-1-thiogalactopyranoside (IPTG) ........224

L

Lactate dehydrogenase (LDH) ........................... 149, 154

Lactic acid......................................................................242

Lactose Dehydrogenase Elevating Virus

(LDEV)........................................................195

Long-term stable productivity .....................................224

M

Mammalian cell culture, bioreactors

antifoaming agents...................................................... 5

automatization and detailed control........................ 11

bioreactors ................................................................... 3

gas transfer........................................................... 13

preparation .............................................................7

setpoints............................................................. 7–8

cell culture media ................................................3–5, 9

cell expansion .............................................................. 7

controllers.................................................................... 5

equipment.................................................................... 3

feeding strategy ......................................................... 10

feeds and supplements ............................................ 3–5

harvest and clarification ............................................ 10

270 ^{BIOREACTORS IN STEM CELL BIOLOGY: METHODS AND PROTOCOLS}

Index