Table 1

Formulation for IF9S medium

Medium component

(stock concentration)

Source

Volume

added

(250 mL final

volume)

Final

concentration

IMDM

Iscove’s modified Dulbecco’s medium

(IMDM), no phenol red (Gibco, cat# 21056-

023)

117.25 mL

–

F12

Ham’s F-12 nutrient mix, GlutaMAX supplement

(Gibco, cat# 31765-027)

117.25 mL

–

PVA (5%)

Polyvinyl alcohol

(Sigma-Aldrich, cat# P8136-250G)

50 μL

10 mg/L

Lipids (100)

Chemically defined lipid concentrates (Gibco,

cat# 11905031)

250 μL

0.1% (vol%)

ITS-X (100)

Insulin–transferrin–selenium–ethanolamine

(Gibco, cat# 51500-056)

5 mL

2% (vol%)

αMTG (1.3% in

IMDM)

Monothioglycerol

(Sigma-Aldrich, cat# M6145-25ML)

750 μL

40 μ/L

AA2P (5 mg/mL)

Sigma-Aldrich, cat. no. A8960

3.2 mL

64 mg/L

GlutaMAX™

(100)

GlutaMAX-1 supplement

(Gibco, cat# 35050-061)

2.5 mL

1% (vol%)

NEAA (100)

MEM nonessential amino acids solution(100)

(Gibico, cat# 11140-035)

2.5 mL

1% (vol%)

Pen-strep

(5000 U/mL)

Gibco, cat# 15070-063

1.25 mL

0.5% (vol%)

Fig. 1 A random positioning machine (RPM). (a) Biaxial drive rotator. (b) Electronic control system. (c) The

T12.5 flasks in biaxial drive rotator were placed into CO2 incubator for cell cultivation

58

Xiaohua Lei et al.