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Tissue Harvest Protocol for the National Human Neural Stem Cell Resource
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(minimally and preferably within 5-8 hours after death):
1. Brain
2. Liver
3. Muscle
4. Skin
5: Others as the specific case dictates

1. Brain

  • Remove the brain, with cerebellum and brainstem intact; cut mid-sagitally and separate the cerebral hemispheres.
  • Fix 1/2 of brain in formalin and store for several weeks; then drain off excess formalin and repackage in a heat-sealed plastic bag (x2) or airtight plastic container and FEDEX to the repository, billing recipient (see notes).
  • Remove the brainstem and cerebellum from the other half of the brain by sectioning through the cerebral peduncles.
  • Section the cerebellum, longitudinally, just lateral to the most lateral extent of the brainstem.
  • Section the remaining hemisphere, placing the medial surface down, coronally at 1cm intervals from frontal to occipital poles.
  • With the remainder of the brain: place each section on a freezing tray (aluminum foil over a block of dry ice or if a block is unavailable, aluminum foil over a stainless steel tray over pellets of dry ice). Cover the sections with powdered dry ice as they are placed.
  • Once the sections are frozen (about 5 minutes), wrap each in the foil and label in the order each section was cut. These foil-wrapped sections can be placed in a ziploc freezer storage bag and stored in the -70° C freezer until they are shipped to the repository (see notes).

2. Liver
Place approximately 1 gram of liver into the same container of fixative as the brain. Freeze a 1cm thick slab of liver in the same manner as the brain.

3. Muscle
Place approximately 1 g of muscle into the same fixative container as the brain. Freeze approximately 10g of muscle in the same manner as the brain (generally, any skeletal muscle will do but it may depend on the clinical history).

4. Skin
Place a 10mmx3mm piece of skin with underlying adipose removed into sterile culture medium in a sterile falcon tube and FEDEX to the repository. If a fibroblast culture exists, this step will not be necessary.


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