Limits...
Human ESC-derived dopamine neurons show similar preclinical efficacy and potency to fetal neurons when grafted in a rat model of Parkinson's disease.

Grealish S, Diguet E, Kirkeby A, Mattsson B, Heuer A, Bramoulle Y, Van Camp N, Perrier AL, Hantraye P, Björklund A, Parmar M - Cell Stem Cell (2014)

Bottom Line: Considerable progress has been made in generating fully functional and transplantable dopamine neurons from human embryonic stem cells (hESCs).We show long-term survival and functionality using clinically relevant MRI and PET imaging techniques and demonstrate efficacy in restoration of motor function with a potency comparable to that seen with human fetal dopamine neurons.Furthermore, we show that hESC-derived dopamine neurons can project sufficiently long distances for use in humans, fully regenerate midbrain-to-forebrain projections, and innervate correct target structures.

View Article: PubMed Central - PubMed

Affiliation: Developmental and Regenerative Neurobiology, Department of Experimental Medical Science, Wallenberg Neuroscience Center, Lund University, 22184 Lund, Sweden; Lund Stem Cell Center, Lund University, 22184 Lund, Sweden. Electronic address: shane.grealish@med.lu.se.

Show MeSH

Related in: MedlinePlus

Functional Equipotency of Intrastriatal Transplanted hESC-DA Neurons and Comparable Morphology to Grafts of Fetal VM-Derived DA Neurons(A) 6-hydroxydopamine lesioned rats displayed a strong unilateral rotational bias upon administration of amphetamine, which was significantly normalized 16 weeks after intrastriatal transplantation of hESC-DA neurons (white bars, n = 5), whereas lesion controls (black bars, n = 6), not receiving any transplant, showed no significant changes when monitored in parallel.(B) Estimation of the number of surviving TH+ neurons within the graft revealed a relatively low number that gave rise to graft-mediated functional recovery.(C) Histological analysis revealed grafts with sparse numbers of TH+ neurons dispersed throughout.(D and E)When comparing transplants of hESC-derived (D) and fetal-derived (E) DA neurons 6 months posttransplantation, both groups revealed grafts rich in DA neurons that are indistinguishable on a gross morphological level.See also Figure S2. TH, tyrosine hydroxylase. In (A), data are represented as mean ± SEM. ∗∗p < 0.01. In (C)–(E), scale bars represent 100 μm.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4232736&req=5

fig2: Functional Equipotency of Intrastriatal Transplanted hESC-DA Neurons and Comparable Morphology to Grafts of Fetal VM-Derived DA Neurons(A) 6-hydroxydopamine lesioned rats displayed a strong unilateral rotational bias upon administration of amphetamine, which was significantly normalized 16 weeks after intrastriatal transplantation of hESC-DA neurons (white bars, n = 5), whereas lesion controls (black bars, n = 6), not receiving any transplant, showed no significant changes when monitored in parallel.(B) Estimation of the number of surviving TH+ neurons within the graft revealed a relatively low number that gave rise to graft-mediated functional recovery.(C) Histological analysis revealed grafts with sparse numbers of TH+ neurons dispersed throughout.(D and E)When comparing transplants of hESC-derived (D) and fetal-derived (E) DA neurons 6 months posttransplantation, both groups revealed grafts rich in DA neurons that are indistinguishable on a gross morphological level.See also Figure S2. TH, tyrosine hydroxylase. In (A), data are represented as mean ± SEM. ∗∗p < 0.01. In (C)–(E), scale bars represent 100 μm.

Mentions: We and others have previously shown that hESC-DA neurons can release DA in vivo (Kirkeby et al., 2012a) and restore a number of motor deficits in 6-OHDA rodent models of PD when assessed 16–18 weeks after grafting (Kirkeby et al., 2012a; Kriks et al., 2011). However, these transplants contained 15,000–18,000 hESC-derived DA neurons, and the high cell number makes it difficult to estimate the efficacy of the cells (Barker, 2014). In previous preclinical and experimental studies using grafts of human fetal VM, it has been shown that normalization of amphetamine-induced rotation after intrastriatal transplantation can be achieved with an average number of 1,200 surviving TH+ neurons (Brundin et al., 1986). On the basis of this information, we performed a potency experiment designed to determine the minimal number of hESC-DA neurons capable of inducing functional recovery in the amphetamine-induced rotation test. We transplanted 6-OHDA lesioned rats, aiming for at least a 10-fold lower number of hESC-DA than the grafts functionally assessed in earlier studies (Kirkeby et al., 2012a; Kriks et al., 2011). The rats were pretested to confirm the completeness of the lesion and immunosuppressed with daily injections of ciclosporin for 18 weeks. At this time point, the rats showed a significant recovery in amphetamine-induced rotation (Figure 2A: t4 = 6.76, p < 0.01; n = 5), despite a much lower number of TH+ neurons in the grafts (Figures 2B and 2C). Quantifications showed that the average number of surviving TH+ neurons was 986 ± 333 per rat (n = 5). Two of the rats had less than 500 surviving hESC-DA neurons and yet showed a complete reduction in rotational bias (Figure 2B). Within TRANSERUO, a EU-funded research consortium formed to develop an efficacious and safe treatment methodology for PD using fetal cell based treatments (http://www.transeuro.org.uk), research groups including our own have optimized and standardized tissue preparation protocols across several centers throughout Europe. A recent report with cells prepared using this protocol show that a significant reduction in amphetamine-induced rotation could be achieved with transplants of human fetal VM with as few 657 ± 199 surviving TH+ neurons (Rath et al., 2013). These results indicate that the functional potency of grafted hESC-DA neurons is on par with that of human DA neurons obtained from fetal VM (Brundin et al., 1986; Rath et al., 2013).


Human ESC-derived dopamine neurons show similar preclinical efficacy and potency to fetal neurons when grafted in a rat model of Parkinson's disease.

Grealish S, Diguet E, Kirkeby A, Mattsson B, Heuer A, Bramoulle Y, Van Camp N, Perrier AL, Hantraye P, Björklund A, Parmar M - Cell Stem Cell (2014)

Functional Equipotency of Intrastriatal Transplanted hESC-DA Neurons and Comparable Morphology to Grafts of Fetal VM-Derived DA Neurons(A) 6-hydroxydopamine lesioned rats displayed a strong unilateral rotational bias upon administration of amphetamine, which was significantly normalized 16 weeks after intrastriatal transplantation of hESC-DA neurons (white bars, n = 5), whereas lesion controls (black bars, n = 6), not receiving any transplant, showed no significant changes when monitored in parallel.(B) Estimation of the number of surviving TH+ neurons within the graft revealed a relatively low number that gave rise to graft-mediated functional recovery.(C) Histological analysis revealed grafts with sparse numbers of TH+ neurons dispersed throughout.(D and E)When comparing transplants of hESC-derived (D) and fetal-derived (E) DA neurons 6 months posttransplantation, both groups revealed grafts rich in DA neurons that are indistinguishable on a gross morphological level.See also Figure S2. TH, tyrosine hydroxylase. In (A), data are represented as mean ± SEM. ∗∗p < 0.01. In (C)–(E), scale bars represent 100 μm.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4232736&req=5

fig2: Functional Equipotency of Intrastriatal Transplanted hESC-DA Neurons and Comparable Morphology to Grafts of Fetal VM-Derived DA Neurons(A) 6-hydroxydopamine lesioned rats displayed a strong unilateral rotational bias upon administration of amphetamine, which was significantly normalized 16 weeks after intrastriatal transplantation of hESC-DA neurons (white bars, n = 5), whereas lesion controls (black bars, n = 6), not receiving any transplant, showed no significant changes when monitored in parallel.(B) Estimation of the number of surviving TH+ neurons within the graft revealed a relatively low number that gave rise to graft-mediated functional recovery.(C) Histological analysis revealed grafts with sparse numbers of TH+ neurons dispersed throughout.(D and E)When comparing transplants of hESC-derived (D) and fetal-derived (E) DA neurons 6 months posttransplantation, both groups revealed grafts rich in DA neurons that are indistinguishable on a gross morphological level.See also Figure S2. TH, tyrosine hydroxylase. In (A), data are represented as mean ± SEM. ∗∗p < 0.01. In (C)–(E), scale bars represent 100 μm.
Mentions: We and others have previously shown that hESC-DA neurons can release DA in vivo (Kirkeby et al., 2012a) and restore a number of motor deficits in 6-OHDA rodent models of PD when assessed 16–18 weeks after grafting (Kirkeby et al., 2012a; Kriks et al., 2011). However, these transplants contained 15,000–18,000 hESC-derived DA neurons, and the high cell number makes it difficult to estimate the efficacy of the cells (Barker, 2014). In previous preclinical and experimental studies using grafts of human fetal VM, it has been shown that normalization of amphetamine-induced rotation after intrastriatal transplantation can be achieved with an average number of 1,200 surviving TH+ neurons (Brundin et al., 1986). On the basis of this information, we performed a potency experiment designed to determine the minimal number of hESC-DA neurons capable of inducing functional recovery in the amphetamine-induced rotation test. We transplanted 6-OHDA lesioned rats, aiming for at least a 10-fold lower number of hESC-DA than the grafts functionally assessed in earlier studies (Kirkeby et al., 2012a; Kriks et al., 2011). The rats were pretested to confirm the completeness of the lesion and immunosuppressed with daily injections of ciclosporin for 18 weeks. At this time point, the rats showed a significant recovery in amphetamine-induced rotation (Figure 2A: t4 = 6.76, p < 0.01; n = 5), despite a much lower number of TH+ neurons in the grafts (Figures 2B and 2C). Quantifications showed that the average number of surviving TH+ neurons was 986 ± 333 per rat (n = 5). Two of the rats had less than 500 surviving hESC-DA neurons and yet showed a complete reduction in rotational bias (Figure 2B). Within TRANSERUO, a EU-funded research consortium formed to develop an efficacious and safe treatment methodology for PD using fetal cell based treatments (http://www.transeuro.org.uk), research groups including our own have optimized and standardized tissue preparation protocols across several centers throughout Europe. A recent report with cells prepared using this protocol show that a significant reduction in amphetamine-induced rotation could be achieved with transplants of human fetal VM with as few 657 ± 199 surviving TH+ neurons (Rath et al., 2013). These results indicate that the functional potency of grafted hESC-DA neurons is on par with that of human DA neurons obtained from fetal VM (Brundin et al., 1986; Rath et al., 2013).

Bottom Line: Considerable progress has been made in generating fully functional and transplantable dopamine neurons from human embryonic stem cells (hESCs).We show long-term survival and functionality using clinically relevant MRI and PET imaging techniques and demonstrate efficacy in restoration of motor function with a potency comparable to that seen with human fetal dopamine neurons.Furthermore, we show that hESC-derived dopamine neurons can project sufficiently long distances for use in humans, fully regenerate midbrain-to-forebrain projections, and innervate correct target structures.

View Article: PubMed Central - PubMed

Affiliation: Developmental and Regenerative Neurobiology, Department of Experimental Medical Science, Wallenberg Neuroscience Center, Lund University, 22184 Lund, Sweden; Lund Stem Cell Center, Lund University, 22184 Lund, Sweden. Electronic address: shane.grealish@med.lu.se.

Show MeSH
Related in: MedlinePlus