medium (1:100) and quickly mix thoroughly by inverting the

tube several times.

4. Plate 3 mL per 6 cm plate or 2 mL per well of a six-well plate.

Store the coated plates at 4^C prior use.

5. Aspirate diluted Geltrex™solution and add pre-warm medium

before use.

2.7

Preparation of

Cell Culture Medium

See Tables 2 and 3.

2.8

Preparation of

Small Molecules

See Table 4.

2.9

Preparation of

Microcarriers

1. Sigmacote a 500-mL glass bottles with Sigmacote^® siliconizing

reagent (Sigma-Aldrich), prevents the microcarriers from

adhering to the glass surface during storage. For coating pro-

tocol, refer to Sigma-Aldrich Product information sheet: VNC,

MAM 10/16-1.

Table 2

500 mL hiPSC culture medium (refer product information sheet, Document #10000003789-PIS_03)

mTeSR™1 Basal medium

400 mL

mTeSR™1 5 supplement

100 mL

Store medium at 2–8 ^C up to 4 weeks

Table 3

1 L Cardiomyocytes (CM) differentiation basal medium (RPMI/B27^-IN)

Medium

Amount added

RPMI-1640 medium (with glucose and L-glutamine)

1 L

50 B-27™supplement, minus insulin

20 mL

L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate

213 mg

Dissolve completely and filter medium using 0.2 μm PES filter

Store medium at 2–8 ^C up to 2 weeks

Table 4

Preparation of small molecules

Small molecule

Stock amount (mg)

Working solution (mM)

Volume of DMSO used (mL)

Y27632 (RI)

10

10

3.122

CHIR99021 (CHIR)

25

12

4.152

Endo IWR1

10

2.5

9.76

Mix well and make 25–50 μL aliquots, to be stored at
20^C

70

Valerie Ho et al.