| Budget Amount *help |
¥3,620,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥420,000)
Fiscal Year 2007: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Research Abstract |
To examine trans-differentiation from human cord blood cells to retinal nerve cells, we used lineage negative fraction from human cord blood. First, low density fraction of human cord blood was obtained using lymphoprep. The low density cells were incubated with biotin-labeled anti-CD3, CD19, CD14, CD15, CD16, CD56, CD11c antibodies, followed by incubation with avidin-labeled magnetic beads. Lineage cells were deleted using a magnet from the low density cells. Residual cells were used as lineage negative cells of human cord blood. The lineage negative cells were labeled with PKH26 and they were injected into subretinal space of SCID mice, in which NK cells had been depleted using anti-asialoGM1 antibody. Two weeks after injection, we sacrificed the mice and obtained the eyes. In histological examination, we detected injected human cells in the eyes of the SCID mice, using a laser-scanning confocal microscope and real time RT-PCR. And the injected cells express human nestin, human MAP2, human neuron specific enolase,β-III tubulin and rhodopsin histologically and they expressed mRNA of human rhodopsin in real time RT-PCR. These results suggest that human cord blood cells can differentiate into retinal nerve cells.
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