Table 1.
Summary of differentiation and transplantation studies of hESCs
| Reference | Differentiation condition | In vitro Characteristics | In vivo Characteristics | ||||||
|---|---|---|---|---|---|---|---|---|---|
| Cell line | Early stage | Maturation stage |
Cell fate | Functional analysis |
Animal model |
Graft innervation | TH+ neuron count/graft |
Behavior improvement |
|
| Perrier et al (2004) | H1, H9, HESC3 | Co-culture with stromal cell | B, AA, G, T, C | TH+/Tuj1+ (64~79%) | DA release Electrophysiology | N/A | N/A | N/A | N/A |
| Ben-Hur et al (2004) | HES1 | Noggin | Neurospheres | TH+/Tuj1+ (0.56%) | N/A | 6-OHDA rat | N/A | 389±83 | APO-IR; stepping and forelimb placing test |
| Buytaert-Hoefen et al (2004) | BG01 | Co-culture with astrocytes/PA6 | Co-culture with G | TH staining | N/A | N/A | N/A | N/A | N/A |
| Schulz et al (2004) | BG01, BG03 | EB, MedII, FGF2 | Neurobasal/B27 FCS, B, G |
TH+/Tuj1+ (75%) TH+/MAP2+ (64%) |
DA release Electrophysiology | 6-OHDA rat | N/A | N/A | N/A |
| Zeng et al (2004) | BG01, variant G01V | Co-culture with PA6 | Co-culture | TH+ (80~90%) | N/A | N/A | N/A | N/A | N/A |
| Park et al (2005) | SNU-hES3, HSF-6, Miz-hES-1 | Co-culture with PA6 and PA6-SHH/FGF8 | N2 or ITSA | TH+/Tuj1+ (41%) | DA release Electrophysiology | 6-OHDA rat | N/A | N/A | No improvement |
| Yan et al (2005) | H9, H1 | EB, SHH, FGF8 | B, AA, G, C | TH+ (30%) | DA release Electrophysiology | N/A | N/A | N/A | N/A |
| Roy et al (2006) | H9, H1 | EB, FGF2, FGF8, SHH, co-culture with hMAST | B, G, FBS | TH+ (54%) | DA release | 6-OHDA rat | N/A | 14,904 ~ 43,234/mm3 | AMP-IR, stepping test, cylinder test |
| Iacovitti et al (2007) | H9, BG01, HUES7, HUES8 | EB, Fibronectin, Noggin, bFGF | C | TH+/Tuj1+ (56~81%) | N/A | 6-OHDA rat | N/A | N/A | N/A |
| Ko et al (2007) | HSF-6, H9 | Co-culture with stromal cell | B, G, C | TH+/Tuj1+ (34~44%) | DA release | 6-OHDA rat | N/A | 201±86 | AMP-IR |
| Sonntag et al (2007) | H7, H9 | Co-culture with MS5-Wnt1, Noggin | B, AA, G, T, C | TH+ (10~25%) | N/A | 6-OHDA rat | N/A | 160±74 | AMP-IR |
| Cho et al (2008) | SNUhES1, SNUhES3, SNUhES16 | EB | Neurospheres, AA, SHH, FGF8 | TH+ (70~80%) En1+/TH+ (91%) DBH+/TH+ (4.1%) PMNT+/TH+ (4.4%) |
DA release Electrophysiology | 6-OHDA rat | N/A | 10,732±4,132 | APO-IR; AMP-IR; stepping test |
| Yang et al (2008) | H9 | SHH, FGF8 | B, AA, G, T, C | TH+/Tuj1+ (43%) | N/A | 6-OHDA rat | TH+/Synaptophysin+ fibers extended out of the graft. | 1,273±340 | AMP-IR |
| Chambers et al (2009) | WA-09 | Noggin and SB B, AA, SHH, FGF8 |
B, AA, G, T, C | TH+/Tuj1+ staining | N/A | N/A | N/A | N/A | N/A |
| Morizane et al (2010) | SA002 | Co-culture with PA6 | B, AA, G, C | TH+/Tuj1+ (17%) | DA release | N/A | N/A | N/A | N/A |
| Cooper et al (2010) | WA-09 | Noggin, RA, Wnt1, FGF8a, B, AA, SHH | B, AA, G, T, C | FOXA2+/TH+/Tuj1+ (5%) | N/A | N/A | N/A | N/A | N/A |
| Kim et al (2010) | H9, Miz-hES4, Miz-hES6, CHA-hES3, SNU-hES3 SNU-hES16 |
EB, SB, DM, SHH, FGF8 | B, AA, G | N/A | N/A | N/A | N/A | N/A | N/A |
| Kriks et al (2011) | H9, H1 | LDN, SB, SHH, FGF8, PMN, CHIR | B, AA, G, T, C, D | TH+ (70%) TH+/FOXA2+ (15~20%) |
DA release Electrophysiology | 6-OHDA mouse, 6-OHDA rat, MPTP monkey | hNCAM+ fibers extended to the non-transplanted brain side | >15,000 | AMP-IR, cylinder test, stepping test |
| Jaeger et al (2011) | H1, H7 | Smad inhibitor, PD0325901, FGF8b, SHH | B, AA, G, T, C | TH+ (25–60%) | Electrophysiology | N/A | N/A | N/A | N/A |
| Morizane et al (2011) | KhES1, 2, 3 | SB, DM, PMN, FGF8 | B, AA, G, C | TH+/Tuj1+ Staining | N/A | N/A | N/A | N/A | N/A |
| Doi et al (2012) | KhES1, 2 | Floating sphere with FGF2 (PA6 conditioned medium, SHH, FGF8) | B, G | TH+/Tuj1+ (12~35%) | DA release | MPTP monkey | N/A | 1.3–18.6×103 | Neurological scores, spontaneous movement |
| Xi et al (2012) | WA09 | SB, LDN, SHH, CHIR, FGF8b | B, AA, G, T, C | TH+ (44%) | Electrophysiology | N/A | N/A | N/A | N/A |
| Kirkeby et al (2012) | H9, SA121 | EB, Noggin, SB, SHH, CHIR | B, AA, G C or D |
FOXA2+/Lmx1a+ (80%) TH staining |
Electrophysiology | 6-OHDA rat | Graft innervated to CPu, Amy, Thal, SN, IC | N//A | AMP-IR, cylinder test |
| Sundberg et al. (2013) | H9 | SB, LDN, SHH, PMN, FGF8a, CHIR, Noggin, SAG, Wnt1, RA | B, AA, G, C, D | TH+ (>10~20%) | N/A | N/A | N/A | N/A | N/A |
| Grealish et al (2014) | H9 | EB, Noggin, SB, SHH and CHIR | B, AA, G, C, D | High co-expression of FOXA2+/Lmx1a+ | N/A | 6-OHDA rat | Graft innervated to CPu, FM, NAc, PL | >1000 | AMP-IR |
| Chen et al. (2016) | H9 | SB, DMH1, SHH, CHIR, SAG, FGF8b | B, AA, G, T, C, C-E | FOXA2+/Lmx1a+ (>80%) TH+ (60%) |
Electrophysiology | 6-OHDA mouse | hNCAM+ fibers innervated to CPu, NAc, Tu | 6,110±254 (61.1±1.0%) | AMP-IR, spontaneous rotation, cylinder test, rota-rod test |
| Kirkeby et al (2017a) | H9, RC17 | SB, Noggin, SHH, CHIR | B, AA, C, D, FGF8b | FOXA2+/Lmx1a+ (90.4±0.9%) | N/A | 6-OHDA rat | Graft innervated to CPu, PL cortex, midbrain | 3,716±1,026 per 100,000 cells grafted |
AMP-IR; cylinder test |
Abbreviations: AA, Ascorbic acid; AMP-IR, Amphetamine-induced rotation; Amy, Amygdala; APO-IR, Apomorphine-induced rotation; B, BDNF; C, Dibutyryl cAMP; C-E, Compound E; CHIR, CHIR99021; CPu, Caudate-putamen; D, DAPT; DM, Dorsomorphin; EB, Embryoid body; FM, Forceps minor; G, GDNF; hNCAM, Human neural cell adhesion molecule; IC, Internal capsule; ITSA, Insulin-Transferrin-Selenium-Sodium Pyruvate; LDN, LDN193189; N/A, Not available; NAc, Nucleus accumbens; PL, Prelimbic cortex; PMN, Purmorphamine; RA, Retinoic acid; SAG, Smo agonist; SB, SB431542; SHH, Sonic Hedgehog; SN, Substantia nigra; T, TGF-β3; Thal, Thalamus; Tu, Olfactory tubercle
Notes: A83-01, TGF β signaling inhibitor; C-E, Notch signaling inhibitor; CHIR, Wnt agonist; D, Notch signaling inhibitor; DM, BMP signaling inhibitor; DMH1, BMP signaling inhibitor; LDN, BMP signaling inhibitor; Noggin, BMP signaling inhibitor; PMN, Shh signaling activator; SB, TGF β signaling inhibitor; SAG, Shh signaling activator