Developing a pure dopamine cell line is a critical step in improving cell replacement
therapies for Parkinson’s. Farber Associate Director Lorraine Iacovitti and colleagues
have shown that it is possible to generate and isolate dopamine neurons from both
mice and human neural stem cells. Here’s what it took to do it:
1. Decode and clone the gene for tyrosine hydroxylase (TH),
an enzyme that is needed for dopamine production by cells.
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2. Insert cloned gene for TH into
fertilized mouse eggs, attached to a gene for Green Fluorescent
Protein (GFP), which causes “tagged” TH cells to glow green.
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2. Insert cloned gene for TH into
human neural stem cells (using a
lentivirus), attached to a gene for
Green Fluorescent Protein (GFP),
which causes “tagged” TH cells
to glow green.
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3. When mice reach embryonic
stage of development,
micro-dissect out the
green-tagged cells to obtain a
high proportion of TH-positive
(dopamine-producing) cells.
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3. Apply growth factors and
other nourishing biochemicals to
induce the cells to differentiate
into dopamine neurons.
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4. Process the cells in a cell-sorting instrument to identify and remove any cells that are not dopamine-specific.
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PURIFIED DOPAMINE CELLS
FOR TRANSPLANTATION |
