Transplanted dopamine neurons derived from primate ES cells preferentially innervate DARPP-32 striatal progenitors within the graft.
The correct identity and functional capacity of transplanted dopamine (DA) neurons derived in vitro from embryonic stem (ES) cells is a critical factor for the development of an ES cell-based replacement therapy for Parkinson's disease. We transplanted primate Cyno-1 ES cells differentiated in vitro for 4 (progenitor ES cells) or 6 (differentiated ES cells) weeks, or control fetal primate cells into the striatum of hemi-parkinsonian rats. Partial behavioral recovery in amphetamine-induced rotation was correlated with the number of ES-derived tyrosine hydroxylase-positive (TH+) neurons in the grafts (r=0.5, P<0.05). Post mortem analysis of ES-derived grafts revealed TH+neurons with mature morphology, similar to fetal DA neurons, and expression of midbrain transcription factors, such as Engrailed (En) and Nurr-1. While the total number of TH+neurons was not different between the two groups, TH/En co-expression was significantly higher (>90%) in grafts from differentiated ES cells than in grafts derived from progenitor cells (<50%), reflecting a more heterogeneous cellular composition. Within the grafts there was an overlap between ES-derived TH+axonal arbors and clusters of primate ES-derived striatal neurons expressing brain factor 1 (Bf-1, Foxg1) and DA and cAMP-regulated phosphoprotein (DARPP-32). Such overlap was never observed for other regional transcription factors that define neighboring forebrain domains in the developing brain, such as Nkx2.1 (medial ganglionic eminence), Nkx2.2 (pallidal and diencephalic progenitors) or Pax6 (dorsal telencephalic progenitors). Despite the heterogeneity of ES-derived graft cell composition, these results demonstrate normal phenotypic specification, conserved natural axonal target selectivity and functionality of DA neurons derived from primate ES cells.
Ferrari
Daniela
D
McLean Hospital/Harvard University Udall Parkinson's Disease Research Center of Excellence, 115 Mill St, Belmont, MA 02478, USA.
Sanchez-Pernaute
Rosario
R
Lee
Hyojin
H
Studer
Lorenz
L
Isacson
Ole
O
D000818:Animals; D001522:Behavior, Animal; D001930:Brain Injuries / Q000139:chemically induced / Q000473:pathology / Q000601:surgery; D002452:Cell Count / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D003342:Corpus Striatum / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D004268:DNA-Binding Proteins / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism*; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32 / Q000378:metabolism*; D005260:Female; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels / Q000378:metabolism; D000090322:Homeobox Protein Nkx-2.2; D007150:Immunohistochemistry / Q000379:methods; D009419:Nerve Tissue Proteins / Q000378:metabolism; D019006:Neural Cell Adhesion Molecules / Q000378:metabolism; D009474:Neurons / Q000502:physiology* / Q000637:transplantation; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D016627:Oxidopamine; D051381:Rats; D017207:Rats, Sprague-Dawley; D033581:Stem Cell Transplantation / Q000379:methods; D013234:Stem Cells / Q000502:physiology*; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism; D019737:Transplants; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D050493:Vesicular Monoamine Transport Proteins / Q000378:metabolism
D004268:DNA-Binding Proteins; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; D000090322:Homeobox Protein Nkx-2.2; C497070:Kcnj6 protein, rat; D009419:Nerve Tissue Proteins; D019006:Neural Cell Adhesion Molecules; C000715628:Nkx2-2 protein, rat; C539884:Nr4a2 protein, rat; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; C493454:Slc18a2 protein, rat; D014157:Transcription Factors; D050493:Vesicular Monoamine Transport Proteins; D016627:Oxidopamine; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine