Table 1.
Summary of studies post-2007 of HLC-derivation from human pluripotent stem cells (adapted from Table 1 and Table 2, Kia et al(13) with modification). Note the limited number of Phase 1 and 2 phenotyping markers generally employed in the characterization of the HLCs.
| Reference | Method of stem cell differentiation | Differentiation efficiency % ALB +ve HLCs | PHENOTYPING: Phase I and II enzyme activity | ||||
|---|---|---|---|---|---|---|---|
| Stem cell (cell line) | Culture format | Differentiation factors | % ALB +ve HLCs (assay method) | Enzyme (assay method) | % hPH comparator | Other comparators | |
| Cai et al., 2007(14) | hESC (H1, H9) | Monolayer, EB formation | AF V, AA, ITS, BMP2, FGF4, HGF, OSM, DEX | 70 (ICC) | CYP2B6 (Fluorescence) | ND | hESC |
| Ek et al., 2007(15) | hESC (SA002, SA002.5, SA167) | Monolayer | Proprietary differentiation medium, FGF2 | ND | CYP1A1 (Fluorescence) | 0 | – |
| CYP3A4 (Fluorescence) | 0 | – | |||||
| Söderdahl et al., 2007(16) | hESC (SA001, SA002, SA002.5, AS034, SA121, and SA167) | Monolayer | Proprietary differentiation medium, bFGF | ND | GST (Fluorescence) | 80 | HepG2 |
| Hay et al., 2008(17); | hESC (H1, H9) | Monolayer | AA, Wnt3a | 90 (ICC) | CYP 1A2 (LC-MS-MS) | 24 | hESC |
| Godoy et al., 2015(11); | CYP1A2 (Luminescence) 100 | ||||||
| Cameron et al., 2015(10) | CYP3A4 (Luminescence) 100 | ||||||
| Shiraki et al., 2008(18) | hESC (Khes-1) | Co-culture with M15 cell line | AA, BMP4, bFGF, HGF, DMSO, DEX, Ly294002 | 9 (ICC) | ND | – | – |
| Agarwal et al.,2008(19) | hESC (WA01, WA09) | Monolayer | AA, FGF4, HGF, BSA, OSM, DEX | 67.4 (ICC) | ND | – | – |
| Moore et al., 2009(20) | hESC (H1) | Monolayer, EB formation | AA, Wnt3a, HGF, OSM, DEX | 72.8 (ICC) | CYP 1A2 (Fluorescence) | ND | hESC-derived HLCs in culture media of different components |
| Basma et al., 2009(21) | hESC (H1) | Monolayer, EB formation | AA, FGF2, HGF, DMSO, DEX | 55.5 (ICC) | CYP1A (Fluorescence) | 30 | – |
| CYP3A (LC-MS-MS) | 90 | – | |||||
| Song et al., 2009(22) | hESC (H1), hiPSC (hFb-derived 3U1, 3U2) | Monolayer | AF V, AA, ITS, BMP2, FGF4, OSM, DEX, KGF, B27 | 60 (ICC) | CYP2B6 (Fluorescence) | ND | hiPSC-derived versus hESC-derived HLCs |
| Duan et al., 2010(23) | hESC (H9) | Monolayer | AA, sodium butyrate, BMP2, BMP4, FGF4, HGF DMSO, B27 | 75-90 (ICC, FACS) | CYP1A2 (LC-MS-MS) | 100 | – |
| CYP2C9 (LC-MS-MS) | 60 | – | |||||
| CYP2D6 (LC-MS-MS) | 95 | – | |||||
| CYP3A4 (LC-MS-MS) | 90 | – | |||||
| Synnergren et al., 2010(24) | hESC (SA002, SA167, SA461) | Monolayer | AA, ITS, FGF1, FGF2, BMP2, BMP4, HGF, OSM, DEX | ND | ND | – | – |
| Touboul et al., 2010(25) | hESC (H9) | Monolayer | AA, BMP4,FGF2, FGF4, FGF10, HGF, EGF, retinoic acid, SB431542, Ly294002 | ND | CYP3A (Bioluminescence) | ND | – |
| Brolén et al., 2010(26) | hESC (SA001, SA002, SA002.5, SA167) | Monolayer | AA, BMP2, BMP4, FGF1, FGF2, HGF, OSM, DEX, Wnt3A | ND | CYP1A (LC-MS-MS) | ND | Spontaneously differentiated hESC-derived HLCs, HepG2 |
| CYP2C (LC-MS-MS) | |||||||
| CYP2A (LC-MS-MS) | |||||||
| Ghodsizadeh et al., 2010(27) | hiPSC (hFb-derived) | EB formation | AA, FGF2, HGF, DMSO, DEX | 50 (FACS) | CYP2B6 (Fluorescence) | ND | hiPSC |
| Liu et al., 2010(28) | hESC (WA01, WA09), hiPSC (hPH-derived) | Monolayer | AA, FGF4, HGF, OSM, DEX | ND | CYP1A2 (Bioluminescence) | ND | – |
| CYP3A4 (Bioluminescence) | |||||||
| Si-Tayeb et al., 2010(29) | hESC (H9), hiPSC (hFb-derived) | Monolayer | AA, BMP4, FGF2, OSM, B27 | 80 (FACS) | ND | – | – |
| Sullivan et al., 2010(30) | hiPSC (hFb-derived) | Monolayer | AA, HGF, Wnt3A, DMSO, OSM, hydrocortisone, tryptose phosphate broth, B27 | 70-90 (ICC) | CYP1A2 (Bioluminescence) | ND | – |
| CYP3A4 (Bioluminescence) | |||||||
| Rashid et al., 2010(31) | hiPSC (hFb-derived) | Monolayer | AA, BMP4, FGF2, HGF, OSM, Ly294002, CHIR99021 (GSK-3 inhibitor) | 83 (FACS) | CYP3A4 (Bioluminescence) | ND | hiPSC |
| Zhang et al., 2011(32) | hESC (H9), hiPSC (hFb-derived) | Monolayer, EB formation | AA, BMP2, FGF4, HGF, KGF, OSM, DEX | 60-80 (ICC, FACS) | CYP3A4 (Bioluminescence) | 0.32 | hESC-derived HLCs |
| Bone et al., 2011(33) | hESC (Shef1, Shef3) | Monolayer | FGF4, HGF, OSM, DEX, 1 m (GSK-3 inhibitor) | ND | ND | – | – |
| Yildirimman et al., 2011(34) | hESC (SA002) | Monolayer | Proprietary differentiation medium | ND | CYP1A2 (LC-MS-MS) | 50 | – |
| CYP3A4 (LC-MS-MS) | 50 | – | |||||
| CYP2B6 (LC-MS-MS) | 10 | – | |||||
| CYP2C9 (LC-MS-MS) | 50 | – | |||||
| CYP2C19 (LC-MS-MS) | 50 | – | |||||
| Chen et al., 2012(35) | hESC (H9), hiPSC (hFb-derived, CFB46) | Monolayer | AA, ITS, HGF, Wnt3A, OSM, DMSO, DEX | ND | CYP3A4 (Bioluminescence) | 100 | hiPSC |
| Cayo et al., 2012(36) | hiPSC (FH patient JD fibroblast-derived) | Monolayer | OCT4, SOX2, NANOG, LIN28 | ND | ND | – | – |
| Schwartz et al., 2012(37) | hiPSC (hFb-derived) | Monolayer | AA, BMP4, FGF2, HGF, OSM | 80 (ICC) | ND | – | – |
| Takayama et al., 2012(38) | hES (H9), hiPSC (hFb-derived, MCR5 & 201B7) | Monolayer | AA, SOX17, HEX, BMP4, FGF4, LacZ, HNF4α, HGF, OSM, DEX | ND | CYP3A4 (Fluorescence) | 100 | – |
| CYP2C9 (Fluorescence) | > 10 | – | |||||
| CYP1A2 (Fluorescence) | < 1 | – | |||||
| Choi et al., 2013(39) | hiPSC (derived from AAT deficient patients) | Monolayer | B27, AA, FGF4, HGF, OSM, DEX | ND | CYP3A4 (Bioluminescence) | 80 | – |
| CYP2D6 (Bioluminescence) | 70 | – | |||||
| CYP2C19 (Bioluminescence) | 90 | – | |||||
| CYP1A2 (Bioluminescence) | 90 | – | |||||
| Ramasamy et al., 2013(40) | hESC (H1) | Monolayer & 3D culture in Algimatrix plate | AA, DMSO, HGF, OSM | ND | CYP3A4 (Bioluminescence) | ND | HepG2 |
| Gieseck et al., 2014(41) | hiPSC (hFb-derived) | Monolayer, 3D-single cell or Clump culture in RAFT system | AA, FGF2,BMP4, LY-294002, Hepatozyme-SFM | ND | CYP3A4 for 2D Day 35 (HPLC-MS) | 4 | – |
| CYP3A4 for 3D Day 45 (Bioluminescence) | 25 | ||||||
| Jia et al., 2014(42) | hiPSC (from urine cells of HA patient) | Monolayer, EB formation | AA, FGF4, BMP2, HGF, KGF, OSM, DEX | 64 (FACS) | ND | – | – |
| Avior et al., 2015(43) | hESC (I3) | Monolayer | AA, B27, Wnt3A, HGF, DMSO, DEX, OSM, FGF2, LCA, MK4 | 83 (FACS) | CYP3A4, 1A2 (Fluorescence) | 30 | HepG2, hESC without LCA/MK4 |
| CYP2E1, 2C9 (Fluorescence) | 8 | ||||||
| Chien et al., 2015(44) | hiPSC (from dental pulp stromal cells) | Co-culture with MEF, EB formation | AF V, AA, FGF4, BMP2, HGF, KFG, OSM, DEX, B27, miR122 (delivered by PU-PEI in CHC) | ND | ND | – | – |
Abbreviations: AA, activin A; AF V, albumin fraction V; bFGF, human recombinant basic FGF; BMP, bone morphogenic protein; BSA, bovine serum albumin; CHC, carboxymethyl-hexanoyl chitosan; DEX, dexamethasone; DMSO, dimethyl sulfoxide; EGF, epidermal growth factor; FACS, fluorescence-activated cell sorting; FGF, fibroblast growth factor; GFP, green fluorescent protein; GSK, glycogen synthase kinase; Hepatozyme-SMF, hepatozyme serum free medium; HEX, hematopoietically-expressed homeobox protein; hESC, human embryonic stem cell; hiPSC, human induced pluripotent stem cell; HLCs, hepatocyte-like cells; HGF, hepatocyte growth factor; HNF4alpha, hepatocyte nuclear factor 4 alpha; ICC, immunocytochemistry; ITS, insulin-transferrin-selenium; KGF, keratinocyte growth factor; LacZ, beta-D-glactosidase; Ly294002, phosphoinositide 3-kinase inhibitor; miR122, microRNA 122; ND, not determined; OCT, octamer-binding transcription factor; OSM, oncostatin M; PU-PEI, biodegradable polyurethane-graft-short-branch polyethylenimine; qRT-PCR, quantitative real time polymerase chain reaction; SB431542, inhibitor for activin receptor-like kinase receptors ALK5, ALK4 and ALK7; SOX, sex determining region Y-box; Wnt3a, wingless-type MMTV integration site family, member 3a