title: Regenerative therapy in experimental parkinsonism: mixed population of differentiated mouse embryonic stem cells, rather than magnetically sorted and enriched dopaminergic cells provide neuroprotection.
abstract: OBJECTIVE
  The objective of the study was to develop regenerative therapy by transplanting varied populations of dopaminergic neurons, differentiated from mouse embryonic stem cells (mES) in the striatum for correcting experimental parkinsonism in rats.
  
  
  METHODS
  mES differentiated by default for 7 days in serum-free media (7D), or by enhanced differentiation of 7D in retinoic acid (7R), or dopaminergic neurons enriched by manual magnetic sorting from 7D (SSEA-) were characterized and transplanted in the ipsilateral striatum of 6-hydroxydopamine-induced hemiparkinsonian rats. Neurochemical, neuronal, glial and neurobehavioral recoveries were examined.
  
  
  RESULTS
  7R and SSEA- contained significantly reduced NANOG and high MAP2 mRNA and protein levels as revealed, respectively, by reverse transcriptase-PCR and immunocytochemistry, compared with 7D. Striatal engraftment of 7D resulted in a significantly better behavioral and neurochemical recovery, as compared to the animals that received either 7R or SSEA-. The 7R transplanted animals showed improvement neither in behavior nor in striatal dopamine level. The grafted striatum revealed increased GFAP staining intensity in 7D and SSEA-, but not in 7R cells transplanted group, suggesting a vital role played by glial cells in the recovery. Substantia nigra ipsilateral to the side of the striatum, which received transplants showed more tyrosine hydroxylase immunostained neurons, as compared to 6-hydroxydopamine-infused animals.
  
  
  CONCLUSIONS
  These results demonstrate that default differentiated mixed population of cells are better than sorted, enriched dopaminergic cells, or cells containing more mature neurons for transplantation recovery in hemiparkinsonian rats.
authors: Division of Cell Biology and Physiology, CSIR-Indian Institute of Chemical Biology, Laboratory of Clinical & Experimental Neuroscience, Jadavpur, India.
authors: Debasmita
authors: D
authors: Tripathy
authors: Poonam
authors: P
authors: Verma
authors: Dominic N
authors: DN
authors: Nthenge-Ngumbau
authors: Meghna
authors: M
authors: Banerjee
authors: Kochupurackal P
authors: KP
authors: Mohanakumar
mesh_terms: D018663:Adrenergic Agents / Q000633:toxicity; D000661:Amphetamine; D000818:Animals; D001058:Apomorphine; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D004195:Disease Models, Animal; D004298:Dopamine / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000502:physiology*; D007839:Functional Laterality / Q000187:drug effects; D008280:Magnetics; D008297:Male; D008474:Medial Forebrain Bundle / Q000293:injuries; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D016627:Oxidopamine / Q000633:toxicity; D020734:Parkinsonian Disorders / Q000139:chemically induced / Q000601:surgery*; D051381:Rats; D017207:Rats, Sprague-Dawley; D033581:Stem Cell Transplantation / Q000379:methods*
chemical_list: D018663:Adrenergic Agents; D009419:Nerve Tissue Proteins; D016627:Oxidopamine; D000661:Amphetamine; D001058:Apomorphine; D004298:Dopamine
keywords: 6-OHDA-induced hemiparkinsonism; Default differentiation; Embryonic stem cells; Intrastriatal transplantation; Manual magnetic cell sorting; Nigral neuronal loss; Retinoic acid