Establishment of the removal method of undifferentiated induced pluripotent stem cells coexisting with chondrocytes using R-17F antibody

Supplementary Figure S1. Evaluation of the appropriate timing to add the secondary antibody The secondary antibody (2nd Ab) was added to co-cultured iPSCs and chondrocytes at the same time as R-17F or 24 h later. GSL-glycan analysis was performed on day 4 of culture. The graph shows the amount of LNFP I. The relative amount of LNFP I according to only R-17F treatment was set to 100%. Error bars indicate the standard deviation of triplicate measurements. Supplementary Figure S2. Side effects of secondary antibody on iPSCs iPSC colonies were treated with the secondary antibody at a concentration of 0–100 μg/mL and observed under a microscope over time. Scale bar = 200 μm. Supplementary Figure S3. Side effects of the secondary antibody on co-cultured cells A) Co-cultured iPSCs and chondrocytes were treated with the secondary antibody at a concentration of 0–100 μg/mL and observed under a microscope over time. Scale bar = 500 μm. B) Quantitative evaluation of iPSCs by GSL-glycan analysis at 72 h after adding the secondary antibody to co-cultured cells. The relative amount of LNFP I without R-17F treatment was set to 100%. Error bars indicate the standard deviation of triplicate measurements. Supplementary Figure S4. LNFP I glycan analysis of transplanted cells A) MALDI-TOF MS spectra of GSL-glycans on a) co-cultured iPSCs and chondrocytes (control), and b) after R-17F and 2nd Ab treatment (+R-17F & 2nd Ab). B) The quantitative amount of LNFP I of control and R-17F and 2nd Ab treatment.