Dissection and Preparation of Human Primary Fetal Ganglionic Eminence Tissue for Research and Clinical Applications
Here, we describe detailed dissection and enzymatic dissociation protocols for the ganglionic eminences from the developing human brain to generate viable quasi-single cell suspensions for subsequent use in transplantation or cell culture. These reliable and reproducible protocols can provide tissue for use in the study of the developing human brain, as well as for the preparation of donor cells for transplantation in Huntington’s disease (HD). For use in the clinic as a therapy for HD, the translation of these protocols from the research laboratory to the GMP suite is described, including modification to reagents used and appropriate monitoring and tissue release criteria.
KeywordsHuntington’s disease Human Dissection Dissociation Fetal tissue Medium spiny neurons Striatum Ganglionic eminence
The protocols described in this chapter were developed using fetal samples from the South Wales Initiative for Fetal Tissue (SWIFT) Research Tissue Bank, and the Cardiff Fetal Tissue Bank (CFTB). These tissue banks have received funding from the Medical Research Council, the Welsh Government, the Brain Repair and Intracranial Neurotherapeutics (BRAIN) Unit funded through Health and Care Research Wales, and EU FP7 projects TransEuro and Repair-HD. The authors would like to acknowledge the help and contributions of all past and present members of the SWIFT team from within the Brain Repair Group, Cardiff University, and University Hospital of Wales.
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