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Additional file 5: Figure S5. Clinical drug K115 could be used as a Rock inhibitor for subculture of pluripotent stem cells. A Day 1 of H9, hUC-iPSC-2, hUC-iPSC-3 passaging by mTeSR1 with Thiazovivin. B Day 4 of H9, hUC-iPSC-2, hUC-iPSC-3 passaging. C Karyotype analysis of H9, hUC-iPSC-2, hUC-iPSC-3 or the 10th generation by Thiazovivin. D Day 1 of H9, hUC-iPSC-2, hUC-iPSC-3 passaging by mTeSR1 with K115. E Day 4 of H9, hUC-iPSC-2, hUC-iPSC-3 passaging. F Karyotype analysis of H9, hUC-iPSC-2, hUC-iPSC-3 for the 10th generation by K115. Scale bar: 100 μm.

Additional file 1: Figure S1. The signaling pathway were reanalyzed between human zygote and hESC. The signaling pathway were reanalyzed between Zygote and ESC P10 from Tang’s single cell RNA-SEQ data. Red color: up-regulation of signaling pathway, Blue color: down-regulation of signaling pathway, fold change > 2, p < 0.05.

Additional file 3: Figure S3. The process of integration-free iPSCs induction from human urine-derived cells. A The process of human urine cells was inducted to be iPSCs. pEP4 EO2S ET2Kand pCEP4-miR302-367were electroporated into human urine cells, and the process of cell shape changing. B The monoclonal hUC-iPSCs by 4I induction medium and karyotype analysis. C The monoclonal hUC-iPSCs by R5 induction medium and karyotype analysis. Scale bar: 100 μm.

Additional file 2: Figure S2. The major cell death pathways of pluripotent stem cell and orbitrap protein analysis. A Pluripotent stem cells after passage for 24 h. B Amplified cell death picture. C Heatmap of orbitrap analysis: from Left to Right: H9, supernatant death cells of H9S, hUC-iPSC-2, supernatant death cells of U2S. D The major cell death ways of pluripotent stem cells. E Venn diagram of up-regulation and down-regulation of protein expression in H9S U2S contrast with H9 and U2.

Additional file 4: Figure S4. DsRed cells in mTeSR1 and OCM175 culture medium and different culture conditions. A H9ds and U2ds O-IPSCs were cultured in mTeSR1. B H9ds and U2ds O-IPSCs were cultured in OCM175. C H9ds and U2ds O-IPSCs were cultured in mTeSR1 and OCM175 without changing medium for 2 days.

Additional file 6: Table S1. Composition of 4I induction Medium” has been changed to” Composition of 4I induction Medium for hUC-iPSCs.