Neuroprotective effect of neural stem cell-conditioned media in in vitro model of Huntington's disease.
Although neural stem cell (NSC) transplantation has been investigated as a promising tool for reconstituting damaged brains, recent evidences suggest that NSCs may rescue the brain via paracrine effects rather than by direct cell replacements. In this study, we attempted to determine the neuroprotective effect of NSC-conditioned media (NSC-CM) in in vitro model of Huntington's disease. Cerebral hybrid neurons (A1) were transfected with either wild-type huntingtin (18 CAG repeats) or mutant huntingtin (100 CAG repeats). At 24h after the transfection, immunocytochemical patterns of the huntingtin aggregations, as well as the level of N-terminal proteolytic cleavages of huntingtin were analyzed. Neuronal apoptosis was evaluated with flowcytometry after Annexin-V and propidium iodide (PI) staining. Cerebral hybrid neurons transfected with mutant huntingtin showed five aggregates patterns, including diffuse cytoplasmic, dispered vacuoles, perinuclear vacuoles, nuclear inclusions (NI), and cytoplasmic inclusions (CI). NSC-CM reduced the levels of nuclear and cytoplasmic inclusions. The transfection with mutant huntingtin increased the level of N-terminal cleavages, which was reduced by the NSC-CM treatment. In addition, NSC-CM reduced the Annexin-V(+)PI(+) and Annexin-V(+)PI(-) neurons which were induced by the mutant huntingtin transfection. In summary, NSC-CM was neuroprotective in in vitro model of Huntington's disease with modulating mutant huntingtin-induced cytotoxicity.
Lim
Heon-Chang
HC
Department of Anatomay, College of Medicine, Seoul National University, Seoul, South Korea.
Lee
Soon-Tae
ST
Chu
Kon
K
Joo
Kyung Min
KM
Kang
Lami
L
Im
Woo-Seok
WS
Park
Joung-Eun
JE
Kim
Seung U
SU
Kim
Manho
M
Cha
Choong-Ik
CI
D017209:Apoptosis / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005333:Fetus; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000737:chemistry / Q000187:drug effects*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009687:Nuclear Proteins / Q000235:genetics; D013234:Stem Cells / Q000737:chemistry*; D013687:Telencephalon / Q000166:cytology; D014162:Transfection / Q000379:methods; D019680:Trinucleotide Repeat Expansion / Q000235:genetics*
D017077:Culture Media, Conditioned; C086055:HTT protein, human; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D018696:Neuroprotective Agents; D009687:Nuclear Proteins