The dialysis culture plate was prepared in the same way as the previous study [17 (link)]. Briefly, the original membrane of a 6-well culture insert (Corning, New York, NY, USA) was replaced with a regenerated cellulose dialysis membrane (Spectrum Labs, San Francisco, CA, USA), which had a 3 kDa molecular weight cut off (MCWO). The dialysis membrane combined insert cup was inserted in the deep well plate (Corning,) (Figure 1a). During the differentiation, 2.5 mL of differentiation medium was added to the culture insert and 14.5 mL of dialysis medium was added in the deep well (Figure 1b). The molecule larger than 3 kDa remained in the culture insert during the culture on an orbital lab shaker. To compare the dialysis culture with a conventional suspension culture without dialysis, the dialysis membrane of the culture insert was sealed with polyethylene film (Diversified Biotech, Dedham, MA, USA) to block the mass transfer by the dialysis.
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