1. Supplementary Materials
Culture Medium Refinement by Dialysis for the Expansion of Human Induced Pluripotent Stem Cells in Suspension Culture
Suman Chandra Nath1, Eiji Nagamori1, Masanobu Horie2, Masahiro Kino-oka1
1 Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, Osaka, , Japan
2 Division of Biochemical Engineering, Radioisotope Research Centre, Kyoto University, Kyoto, , Japan
Corresponding author
Masahiro Kino-oka
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2. Supplementary Fig. S1 Effect of lactic acid on hiPSC growth
Supplementary Fig. S1 Effect of lactic acid on hiPSC growth. Growth rate of hiPSC at different concentration of lactic acid in adherent culture. *p < 0.01.

3. Supplementary Fig. S2 Different storage conditions of fresh medium for checking bFGF aggregation, stabilization with heparin and consumption by cells after 24 h at 37°C. *p < 0.01.

4. Supplementary Table S1 Flow cytometry analysis for OCT 3/4 and TRA-1-60 during the determination of lactic acid toxicity limit, adherent culture and suspension culture
OCT 3/4 (%)
TRA-1-60 (%)
Lactic acid toxicity limit
0 g L-1
99.5 ± 0.3
62.6 ± 6.0
0.5 g L-1
99.1 ± 0.2
53.8 ± 4.1
1.0 g L-1
94.5 ± 2.6
50.1 ± 3.0*
Adherent culture
Fresh medium
99.2 ± 0.6
68.4 ± 4.5
Refined+bFGF
98.6 ± 0.4
71.6 ± 4.0
Refined
94.7 ± 3.5
47.4 ± 3.8*
Suspension culture
98.1 ± 0.9
69.2 ± 2.1
Refining system
99.5 ± 0.2
78.3 ± 1.1*
*p < 0.01

5. Supplementary Fig. S3 Remaining amount of cytokines in the used culture medium. Concentration of (a) bFGF and (b) TGF-β1 during culture with fresh medium (open bar) and medium refining system (close bar). * p < 0.05.

6. Supplementary Table S2 Comparison of medium components between refined and fresh (mTeSR™1) medium
Refined medium
Fresh (mTeSR™1) medium
DMEM-F12 √
L-Ascorbic acid -
Selenium
Transferrin *
NaHCO3
Glutathione
L-Glutamine
Defined lipids
Thiamine
Trace element B
Trace element C
BSA
bFGF
TGF-β1
Insulin
Pipecolic acid
LiCl
GABA
Autocrine factors
√ : Included in medium
* : Expected to be contained in the refined medium
- : Expected not to be contained in the refined medium
Blank: Not included in medium