The Journal of Neuroscience, November 29, 2006, ():
Transplantation of Human Neural Stem Cells Exerts Neuroprotection in a Rat Model of Parkinson's Disease
J. Neurosci. Yasuhara et al.
26: 12497
Supplemental Data
Files in this Data Supplement:
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Supplementary Figure 1: Experimental protocols. A: In vivo study design. BrdU was injected once a day for 2 weeks before sacrifice. In addition to the vehicle injection, single SCF/Culture media (CM) injection, continuous SCF infusion, or dead GFP-HB1.F3 cell transplantation was performed as controls. Furthermore, rats were sacrificed at the earlier time points (3 days, 1 week, and 2 weeks post-transplantation) in order to reveal the surviving rate of the transplanted GFP-HB1.F3 cells. B: 0, 1, 10 or 40μM of 6-OHDA was added to SH-SY5Y cell culture and after 12 or 24 hours, cell viability was evaluated by MTT assay or immunocytochemically. C: Culture media (CM) from HB1.F3 cells was administered at the same time as 6-OHDA was delivered to SH-SY5Y neurons. Neuroprotective effects of CM were examined after 24 hours. D: HB1.F3 cells were initially cultured for 24 hours, and thereafter fixed. SH-SY5Y cells were grown on fixed HB1.F3 cells for 48 hours, then exposed to 6-OHDA for 24 hours, and cell viability assessed thereafter. E: Similar to paradigm C, except that SCF and/or blocking antibodies, in addition to CM and 6-OHDA, were administered in each experiment. F: TUNEL stain was performed at 12 hours after CM treatment. G: Similar to paradigm B, except that c-kit expression was assessed at 24 hours after 6-OHDA exposure. H: For Bcl-2 ELISA, cell lysates were collected at specific time points after drug treatment. I: Neuroprotective effects of SCF and CM on 6-OHDA-exposed dopaminergic neurons from fetal rat ventral mesencephalon were explored. After culture for 96 hours, the same paradigm was used for cell viability and c-kit expression.
