Table 1.
Summary of preclinical stem cell studies in amyotrophic lateral sclerosis
| Reference | Donor spec. | Cell type | Modification | Experimental model | Delivery method | Treatment age | Adjunct therapy | Functional improvement | Survival benefit | Histologic change | Other comments |
|---|---|---|---|---|---|---|---|---|---|---|---|
| Lopez-Gonzales (2009) [11] | Mouse | ES | MN differentiated in vitro; GFP expressing | SOD1-G93A rat | 1 × 10^5 cells, intraspinal | 10 weeks | Ciclosporin A | Delay in motor decline by Rotarod | None | n/a | Degeneration of grafted cells at endstage |
| Deshpande (2006) [15] | Mouse | ES | MN differentiated in vitro; GFP expressing | Neuroadapted Sindbis virus-induced motor neuron death, rat | 6 × 10^4 cells, intraspinal | 5–7 weeks | Ciclosporin A; dibutyryl cyclic adenosine monophosphate, GDNF and/or rolipram | Recovery of grip strength in animals receiving all supplemental treatments | n/a | n/a | n/a |
| Harper (2004) [16] | Mouse | ES | MN differentiated in vitro; GFP expressing | Neuroadapted Sindbis virus-induced motor neuron death, rat | 6 × 10^4 cells, intraspinal | 5–7 weeks | Ciclosporin A; Rho kinase inhibitor (Y27632) or dibutyryl cyclic adenosine monophosphate | n/a | n/a | n/a | Axonal growth facilitated by dibutyryl cyclic adenosine monophosphate |
| Kerr (2003) [17] | Human | ES | none | Neuroadapted Sindbis virus-induced motor neuron death, rat | 3 × 10^5 cells, intrathecal | 3–4 weeks | Ciclosporin A or FK-506 | Recovery of motor function by BBB and grip strength testing | n/a | Increased motor neuron survival | Possibly mediated via TGF-α or BDNF |
| Corti (2004) [18] | Mouse | BM | None | SOD1-G93A mouse | 3 × 10^7 cells intraperitoneal | 4 weeks | Irradiation, 800 rad | Delay in motor decline by Rotarod | 10–13 days | Increased motor neurons and ventral root axons at 100 days | A large proportion of microglial cells formed |
| Solomon (2006) [19] | Mouse | BM | GFP expressing | SOD1-G93A mouse | 5 × 10^6 cells, intravenous | 6 weeks | Irradiation, 950–1100 rads | None | None | Large proportion of transplanted cells with microglial markers integrated into spinal cord | n/a |
| Ohnishi (2009) [20] | Mouse | BM | GFP expressing | SOD1-G93A mouse | 6 × 10^7 cells intra-bone marrow | 87.7–89 days | Irradiation, 6 Gy × 2 doses | Delay in motor decline by grip strength meter | 10–13 days | Increased motor neurons and ventral root axons in both eGFP and mutant SOD1 BM transplanted animals over wild type | Microglial markers noted in transplanted cells |
| Pastor (2013) [21] | Mouse | BM | GFP expressing or GDNF knockout | mdf mouse | 1 × 10^6 cells, intramuscular | 10 weeks | None | Improvement in motor function by rotarod and treadmill testing | n/a | Improvement in neuromuscular junction and muscle histology; increased motor neuron survival on the side of treated limb | Increased GDNF expression noted in spinal cord and corresponding cortex |
| Cabanes (2007) [22] | Mouse | BM | CD117+, GFP expressing | mdf mouse | 3 × 10^5 cells, intraspinal | 6 weeks | n/a | Improvement by footprint testing | n/a | Increase motor neuron survival | GDNF levels higher in grafted mice |
| Corti (2010) [23] | Mouse | BM | Lin-c-kit+ | SOD1-G93A mouse | 1 × 10^6 cells intravenous | 70 days | n/a | Delay in motor decline by Rotarod | 16–17 days | Increased motor neuron survival (~50 %) with preserved ventral nerve root axons | Mechanism of protection possibly mediated by expression of GLT1 and elaboration of VEGF and angiopoietin 2 |
| Pastor (2012) [24] | Mouse | BM, MSC | GFP expressing or GDNF knockout | mdf mouse | 0.5–1 × 10^6 cells, intraspinal | n/a | None | Greater improvements seen in BM transplants over MSC transplants | n/a | Transplanted BM proliferate and retain bone marrow phenotype; MSCs underwent apoptosis | Increased GDNF expression in BM transplanted animals, functional improvements abolished in GDNF knockout BM transplant |
| Garbuzova-Davis (2008) [25] | Human | UCB | none | SOD1-G93A mouse | 1–5 × 10^7 cells intravenous | 7–8 weeks | Ciclosporin A | Delay and improvement in hindlimb extension and Rotarod | ~13 days (2.5 × 10^7 cell group) | Reduced microglial density in spinal cord | n/a |
| Garbuzova-Davis (2003) [26] | Human | UCB | None | SOD1-G93A mouse | 1 × 10^6 cells intravenous | 9 weeks | Ciclosporin A | Delay in motor decline by extension reflex and footprint testing | None | n/a | Distributed throughout CNS and formed cells with astrocyte and neuronal markers |
| Souayah (2012) [27] | Human | UCB | None | SOD1-G93A mouse | 1 × 10^8 cells, intravenous | 5–6 weeks | None | n/a | n/a | n/a | Improved neuromuscular transmission by electrodiagnostic testing |
| Bigini (2011) [28] | Human | UCB | None | SOD1-G93A mouse; Wobbler mouse | 5 × 10^5 cells, intraventricular | 10 weeks (SOD1); 4 weeks (Wobbler) | Ciclosporin A | Slowed decline by stride length and rotarod (SOD1); slowed decline by running speed and grip strength (Wobbler) | 18 days | No difference in MN (SOD1); increased MN survival (Wobbler) | n/a |
| Knippenberg (2012) [29] | Human | UCB | CD34+ | SOD1-G93A mouse | 2 × 10^5 cells, intraspinal | 40 or 90 days | Ciclosporin A | Improvement in motor function by rotarod, stride length, and footprint analysis | 6 days (12 days in females) | Increased motor neuron survival (~50 %) | No detected changes in growth factor production |
| Rizvanov (2011) [30] | Human | UCB | VEGF and FGF2 expressing | SOD1-G93A mouse | 1 × 10^6 cells intravenous | 24–28 weeks | n/a | n/a | n/a | n/a | VEGF-FGF2 expressing cells demonstrated astrocyte markers |
| Rizvanov (2008) [31] | Human | UCB | VEGF and L1CAM expressing | SOD1-G93A mouse | 1 × 10^6 cells, intravenous | 22–25 weeks | None | n/a | n/a | Transplanted cells formed endothelial cells | n/a |
| Habisch (2007) [32] | Human | MSC, UCB | Neuroectodermal derivatives of each also used | SOD1-G93A mouse | 1 × 10^5 cells, intrathecal | 45 days | Ciclosporin A | None | None | Improved intraparenchymal incorporation with bone marrow-derived cells | n/a |
| Uccelli (2012) [33] | Mouse | MSC | Luciferase expressing | SOD1-G93A mouse | 1 × 10^6 cells, intravenous | 90 days | None | Improvement by rotarod, extension reflex, and motor score | 17 days | Reduced ubiquitin inclusions | Decreased activated astrocyte and microglial cells, improvement in profile of oxidative stress/antioxidant enzyme expression |
| Forostyak (2011) [34] | Rat | MSC | GFP expressing | SOD1-G93A rat | 1 × 10^5 cells intraspinal, 2 × 10^6 intravenous | 16 weeks | Ciclosporin A | Slowed decline in motor function by BBB and grip strength testing | 11 days | Increased motor neuron survival in treated group | n/a |
| Boucherie (2009) [35] | Rat | MSC | None | SOD1-G93A rat | 2 × 10^6 cells, intrathecal | 90 days | None | Delayed disease onset by motor score | 16 days | Increased motor neuron survival and significant proportion formed astrocyte-like cells, with decreased microglia | Reduced expression of inflammatory mediators |
| Zhao (2007) [36] | Human | MSC | None | SOD1-G93A mouse | 3 × 10^6 cells, intravenous | 8 weeks | Irradiation, 6 Gy | Improved motor function by rotarod testing | 18 days | Increased motor neuron survival at 16 and 20 weeks, improved CMAP amplitudes | n/a |
| Vercelli (2008) [37] | Human | MSC | None | SOD1-G93A mouse | 1 × 10^5 cells, intraspinal | 28 weeks | none | Improved motor function by rotarod testing | n/a | Increased motor neuron survival and decreased microglial activation | Motor improvement noted in male animals only |
| Kim (2010) [38] | Human | MSC | None | SOD1-G93A mouse | 0.1, 2.0, and 10 × 10^5 cells, intrathecal | 60 days | Ciclosporin A | Slowed decline by rotarod testing (10 × 10^5 cell group) | ~6–8 days (2 × 10^5 and 10 × 10^5 cell groups) | Increased motor neuron survival | n/a |
| Suzuki (2008) [39] | Human | MSC | GFP, GDNF expressing | SOD1-G93A rat | 3.6 × 10^5 cells, intramuscular split in 3 doses | 80 days | Bupivacaine; ciclosporin A | Delay in motor dysfunction by BBB rating | 18–28 days | Preservation of neuromuscular junctions and corresponding motor neurons | n/a |
| Knippenberg (2012) [40] | Human | MSC | Glucagon-like peptide 1 expressing, alginate matrix embedded | SOD1-G93A mouse | 30 alginate capsules | 40 days | None | Improvement in motor function by rotarod and footprint analysis | 13 days | No difference in MN, reduction in reactive astrocytes and microglia | Increased expression of heat shock protein 70 in treated mice |
| Choi (2010) [41] | Human | MSC | Retroviral transduction of Neurogenin 1 | SOD1-G93A mouse | 1 × 10^6 cells intravenous | 8, 14–16, or 13 and 15 weeks | Ciclosporin A | Delay in motor decline by rotarod | n/a | Improved motor neurons survival | n/a |
| Morita (2008) [42] | Mouse/Rat | OEC, MSC | GFP expressing | Leu126delTT mouse | 3 − 4 × 10^5 cells, intraventricular | 13–14 weeks | FK-506 | None | Difference seen only in MSC-treated females | n/a | n/a |
| Martin (2007) [43] | Mouse | OEC | GFP expressing | SOD1-G93A mouse | 0.4 − 120 × 10^5 cells, intraspinal | 70 days | None | Improved motor function by running wheel activity and inclined plane test | 22.3 days | Differentiated cells show markers for MNs, astrocytes, and oligodendrocytes | Axons seen to project to peripheral nerve but fail to make neuromuscular junctions |
| Lepore (2008) [44] | Rat | Glial restricted precursors | GLT1-overexpressing, also GLT1-null cells | SOD1-G93A rat | 9 × 10^5 cells, intraspinal | 90 days | Ciclosporin A | Delay in forelimb (site of transplantation) but not hindlimb function; slowed decline of phrenic nerve peak CMAP amplitudes | 16.9 days | Increased motor neuron survival (47 % increase) | Majority astrocyte differentiation, may be mediated by GLT1 expression |
| Lepore (2011) [45] | Human | Glial restricted precursors | None | SOD1-G93A mouse | 2 − 6 × 10^5 cells, intraspinal | 50–60 days | Ciclosporin A or FK-506/rapamycin | None | None | No difference | n/a |
| Corti (2007) [46] | Mouse | Primary NPC | GFP expressing; LewisX + −CXCR4+ | SOD1-G93A mouse | 2 × 10^4 cells intraspinal | 70 days | n/a | Motor decline delayed, no change in slope of disease progression | 22–23 days | Increased MN survival and preservation of ventral nerve roots at 110 days but not at end stage | A large proportion of neuronal cells formed, with a subset of motor neurons. Neuroprotection may be mediated by VEGF and IGF-1 |
| Mitrecic (2010) [47] | Rat | Primary NPC | GFP expressing | SOD1-G93A rat | 1 × 10^7 cells intravenous | 14 and 26 weeks | Nitroprusside; TNF in half of treatment group | n/a | n/a | n/a | Improvement in graft survival with TNF therapy at 7 days; dominant GFAP positive cells at 7 days |
| Hwang (2009) [48] | Human | NPC | VEGF expressing | SOD1-G93A mouse | 1 × 10^5 cells, intrathecal | 70 days | n/a | Delay in motor decline by rotarod, paw grip endurance and extension reflex testing | 12 days | n/a | Downregulation of apoptotic proteins in treated animals |
| Park (2009) [49] | Human | NPC | GFP with GDNF or multiple growth factors (BDNF, IGF-1, VEGF, NT-3, GDNF) expressing | SOD1-G93A mouse | Intrathecal | 75 days | Ciclosporin A | none | Survival decreased in treatment group | Increased motor neuron survival with GDNF expressing cells | Sexual dimorphism noted, with female animals with worse outcomes |
| Xu (2006) [50] | Human | NPC | None | SOD1-G93A rat | 4 × 10^5 cells, intraspinal | 62 day | FK-506 | Slowed progression by BBB tests | 11 days | Increased motor neuron survival (nearly 200 %) | Increased GDNF secretion in grafted animals |
| Xu (2009) [51] | Human | NPC | none | SOD1-G93A rat | 1.6 × 10^5 cells, intraspinal | 56 day | FK-506 | n/a | n/a | n/a | n/a |
| Yan (2006) [52] | Human | NPC | None | SOD1-G93A mouse | 8 × 10^5 cells, intraspinal | 8 weeks | FK-506 and/or rapamycin and/or mycophenolate mofetil; or anti CD4 antibodies | Improved motor function by modified Wrathall scale with combination immunosuppressants | ~3 week benefit with combination immunosuppressants | Prominently neuronal phenotype formed | Combination immunosuppression promotes survival of grafted cells, which imparts functional and survival benefit |
| Klein (2005) [53] | Human | NPC | GDNF expressing | SOD1-G93A rat | 4.8 × 10^5 cells, intraspinal | 90 days | Ciclosporin A | None | None | None | n/a |
| Suzuki (2007) [54] | Human | NPC | GDNF expressing | SOD1-G93A rat | 4.8 − 7.2 × 10^5 cells, intraspinal | 65 days | Ciclosporin A | none | none | Increased motor neuron survival by grafted cells; no change in neuromuscular junctions | n/a |
| Garbuzova-Davis (2002) [55] | Human | NPC | None | SOD1-G93A mouse | 7.5 × 10^4 cells, intraspinal | 53 days | Ciclosporin A | Suggestion of delayed motor decline by staircase testing, beam balance, open field and footprint testing | None | None | n/a |
| Gao (2005) [56] | Human | NPC | MN differentiated in vitro; GFP expressing | Neonatal sciatic axotomy rat | 1 × 10^5 cells, intraspinal | 2 months | Ciclosporin A | Improvement by sciatic function index | n/a | 51 % transplanted cells with MN markers, and 19 % taking up retrograde marker from muscle | n/a |
| Hwang (2009) [48] | Human | NPC | VEGF expressing | SOD1-G93A mouse | 1 × 10^5 cells, intrathecal | 70 days | None | Delayed disease onset by rotarod, paw grip endurance and extension reflex testing | 12 days | Some transplanted cells differentiated into MN-like cells | Reduced proapoptotic proteins and elevated antiapoptotic proteins |
| Xu (2011) [57] | Human | NPC | None | SOD1-G93A rat | 2.4 × 10^5 cells, intraspinal | 63 days | FK-506 | Delayed motor decline by BBB and inclined plane testing | 17 days | n/a | n/a |
| Hefferan (2012) [58] | Human | NPC | None | SOD1-G93A rat | 1.8 − 2.6 × 10^5 cells, intraspinal | 60–65 days | FK-506 and mycophenolate mofetil | Improvement in motor function by BBB scale and electrodiagnostic testing | None | Increased motor neuron survival (~43 %); decreased reactive astrocyte and microglial populations | n/a |
| Popescu (2013) [59] | Human | iPSC-NPC | None | SOD1-G93A rat | 1 × 10^5 cells, intraspinal | 3 months | Ciclosporin A | n/a | n/a | Cells formed neuronal phenotype, with MN-like morphology | n/a |
| Nizzardo (2014) [60] | Human | iPSC-NPC | ALDHhi-SSClo-VLA4+; GFP expressing | SOD1-G93A mouse | 1 × 10^6 cells per dose, intravenous (multiple doses) or intrathecal (3 doses) | 90 days | None | Improvement in motor function by rotarod | 10 days (i.t.); 23 days (i.v.) | Increased motor neuron survival (40 %) and preservation of ventral axons (50 %) | Decreased microglial activation and astrogliosis |
ALDH aldehyde dehydrogenase, BBB Basso-Beatti-Bresnahan scale, BDNF brain derived neurotrophic factor, BM bone marrow, CMAP compound muscle action potential, ES embryonic stem cell, FGF fibroblast growth factor, GDNF glial cell-line derived neurotrophic factor, GFP green fluorescent protein, GLT1 glutamate transporter 1, IGF-1 insulin-like growth factor 1, iPSC induced pluripotent stem cell, L1CAM L1 cell adhesion molecule, MN motor neuron, MSC mesenchymal stem cells, NPC neural progenitor cell, NT-3 neurotrophin 3, OEC olfactory ensheathing cell, SSC side scatter, TGF transforming growth factor, UCB umbilical cord blood cells, VEGF vascular endothelial growth factor, VLA4 very late antigen 4 (integrin alpha 4 beta 1) SOD1 superoxide dismutase; n/a not applicable; CNS central nervous system; i.t intrathecal; i.v. intravenous