Table S1. Materials used in hESCs culture and differentiation.

Material / Company / Cat. Number
Matrigel / Sigma-Aldrich / E1270
Neurobasal / Invitrogen / 21103049
DMEM/F12 / Invitrogen / 11320082
MEM non-essential amino acid / Invitrogen / 11140050
L-Glutamin / Invitrogen / 25030081
Knock out serum replacement / Invitrogen / 12618-013
DPBS / Invitrogen / 14287-072
PBS, without Ca2+, Mg2+ / Invitrogen / 21600-010
FBS / Thermo Scientific Hyclone / SH30071
RPMI 1640 / Invitrogen / 51800
Insulin-transferrin-selenium / Invitrogen / 41400-045
Y-27632 dihydrochloride monohydrate / Sigma-Aldrich / Y0503
Activin A / R&D Systems / 338-AC
Fibroblast growth factor / R&D Systems / 235-F4
Hepatocyte growth factor / R&D Systems / 294-HG
Oncostatin M / R&D Systems / 295-OM
Dexamethasone / Sigma Aldrich / D2915
HCMBulletKit / Lonza / CC-3189
Trypsin-EDTA 0.5% / Invitrogen / 25300054
Collagenase type IV / Invitrogen / 17104-019
Dispase / Invitrogen / 17105-041
DMEM / Invitrogen / 12800-082
Trypsin 1:250 / Invitrogen / 27250-018

TableS2. Materials used in molecular analysis.

Material / Company / Cat. number
TRIZOL / Sigma-Aldrich / T9424
cDNA synthesis kit / Fermentase / K1632
Recombinant DNaseI (RNnase free) / Takara / 2270A
Real Time PCR Kit / ABI / AB4367659

Table S3.Primer sequences.

Gene name / Sequence
SOX17 / F: 5' CTC TGC CTC CTC CAC GAA 3'
R: 5' CAG AAT CCA GAC CTG CAC AA 3'
FOXA2 / F: 5' ATGCACTCGGCTTCCAGTAT 3'
R: 5' TGTTGCTCACGGAGGAGTAG 3'
GATA4 / F: 5' CCT GTC ATC TCA CTA CGG 3'
R:5'GCT GTT CCA AGA GTC CTG 3'
GOOSECOID / F: 5' CAC CAT CTT CAC TGA CGA G 3'
R: 5' GTT CTT AAA CAA GAC CTC CA 3'
NANOG / F: 5' AAA GAA TCT TCA CCT ATG CC 3'
R: 5' GAA GGA AGA GGA GAG ACA GT 3'
BRACHYURY / F: 5' AAT TGG TCC AGC CTT GGA AT 3'
R: 5' CGT TGC TCA CAG ACC ACA 3'
SOX1 / F: 5' CAC AAC TCG GAG ATC AGC AA 3'
R: 5'GGT ACT TGT AAT CCG GGT GC 3'
AFP / F: 5' AAA TGC GTT TCT CGT TGC TT 3'
R: 5' GCC ACA GGC CAA TAG TTT GT 3'
ALB / F: 5' CTT CCT GGG CAT GTT TTT GT 3'
R: 5' TGG CAT AGC ATT CAT GAG GA 3'
HNF4A / F: 5' CGATGACAATGAGTATGCCT 3'
R: 5'GTCGTTGATGTAGTCCTCCA 3'
CYP3A7 / F: 5'GACCGTAAGTGGAGCCTGATTTC3'
R: 5'ACAGACCATGAGAGAGCACAA3'
CYP3A4 / F: 5'TTTTTGGATCCATTCTTTCTCTCAA3'
R: 5'ATCCACTCGGTGCTTTTGTG3'
GAPDH / F: 5' CTC ATT TCC TGG TAT GAC AAC GA 3'
R: 5' CTT CCT CTT GTG CTC TTG CT 3'

TableS4. List of antibodies used in immunoflorescent staining and flowcytometry.

Material / Company / Cat. number / Dilution
Anti-Sox17 antibody / R&D Systems / AF1924 / 1:200
Anti-AFP antibody / Santa Cruz biotechnology / Sc-8108 / 1:100
Anti-ALB antibody / Santa Cruz biotechnology / Sc-51515 / 1:100
Anti-CK18 / Chemicon / MAB1600 / 1:200
Anti-FOXA2 antibody / Abcam / Ab60721 / 1:400
Anti-Goat FITC / Abcam / ab6785 / 1:400
Anti-mouse FITC / Abcam / Ab6785-1 / 1:300
Anti-ASGPR / Santa Cruz biotechnology / Sc-13467 / 1:100
Alexa Flur®488 goat anti mouse / Life Technologies / A11001 / 1:800
Alexa Flur®594 rabbit anti goat / Life Technologies / A21223 / 1:800
Alexa Flur®594 goat anti mouse / Life Technologies / A11020 / 1:800
Bovine serum albumin / Sigma-Aldrich / A3311-100G / 1%
DAPI / Sigma-Aldrich / D8417 / 0.5 µg/ml

Supplementary Fig. 1.Negative controls for immunofluorescence and immunohistofluorescence stainings. Immunofluorescence and immunohistofluorescence stainings were performed on human dermal fibroblasts and untransplanted mouse liver tissues, respectively.

1