Abstract
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.
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References
Walker FO (2007) Huntington’s disease. Lancet 369(9557):218–228
O’Rahilly RR, Müller F (2006) The embryonic human brain: an atlas of developmental stages. John Wiley & Sons, Hoboken, NJ
Grasbon-Frodl E, Nakao N, Lindvall O, Brundin P (1996) Phenotypic development of the human embryonic striatal primordium: a study of cultured and grafted neurons from the lateral and medial ganglionic eminences. Neuroscience 73:171–183
Nakao N, Itakura T (2000) Fetal tissue transplants in animal models of Huntington’s disease: the effects on damaged neuronal circuitry and behavioral deficits. Prog Neurobiol 61:313–338
Hauser RA, Furtado S, Cimino CR et al (2002) Bilateral human fetal striatal transplantation in Huntington’s disease. Neurology 58:687–695
Rosser AE, Barker RA, Harrower T et al (2002) Unilateral transplantation of human primary fetal tissue in four patients with Huntington’s disease: NEST-UK safety report ISRCTN no 36485475. J Neurol Neurosurg Psychiatry 73:678–685
Bachoud-Lévi A-C (2009) Neural grafts in Huntington’s disease: viability after 10 years. Lancet Neurol 8:979–981
Bachoud-Levi AC, Bourdet C, Brugieres P et al (2000) Safety and tolerability assessment of intrastriatal neural allografts in five patients with Huntington’s disease. Exp Neurol 161:194–202
Bachoud-Lévi A-C, Gaura V, Brugières P et al (2006) Effect of fetal neural transplants in patients with Huntington’s disease 6 years after surgery: a long-term follow-up study. Lancet Neurol 5:303–309
Bachoud-Lévi A-C, Rémy P, Nǵuyen JP et al (2000) Motor and cognitive improvements in patients with Huntington’s disease after neural transplantation. Lancet 356(9246):1975–1979
Rosser AE, Bachoud-Lévi A-C (2012) Clinical trials of neural transplantation in Huntington’s disease. Prog Brain Res 200:345–371
Lelos MJ, Roberton VH, Vinh NN et al (2016) Direct comparison of rat- and human-derived ganglionic eminence tissue grafts on motor function. Cell Transplant 25:665–675
Arber C, Precious SV, Cambray S et al (2015) Activin A directs striatal projection neuron differentiation of human pluripotent stem cells. Development 142:1375–1386
Delli Carri A, Onorati M, Lelos M et al (2013) Developmentally coordinated extrinsic signals drive human pluripotent stem cell differentiation toward authentic DARPP-32+ medium-sized spiny neurons. Development 140:301–312
Ma L, Hu B, Liu Y et al (2012) Human embryonic stem cell-derived GABA neurons correct locomotion deficits in quinolinic acid-lesioned mice. Cell Stem Cell 10:455–464
Aubry L, Bugi A, Lefort N et al (2008) Striatal progenitors derived from human ES cells mature into DARPP32 neurons in vitro and in quinolinic acid-lesioned rats. Proc Natl Acad Sci U S A 105:16707–16712
Parmar M, Takahashi J, Studer L, Barker RA (2017) GFORCE-PD still going strong in 2016. NPJ Parkinsons Dis 3:16014
Onorati M, Castiglioni V, Biasci D et al (2014) Molecular and functional definition of the developing human striatum. Nat Neurosci 17:1804–1815
Straccia M, Carrere J, Rosser AE, Canals JM (2016) Human t-DARPP is induced during striatal development. Neuroscience 333:320–330
Straccia M, Garcia-Diaz Barriga G, Sanders P et al (2015) Quantitative high-throughput gene expression profiling of human striatal development to screen stem cell-derived medium spiny neurons. Mol Ther Methods Clin Dev 2:15030
Precious SV, Kelly CM, Reddington AE et al (2016) FoxP1 marks medium spiny neurons from precursors to maturity and is required for their differentiation. Exp Neurol 282:9–18
Schmidt RH, Björklund A, Stenevi U (1981) Intracerebral grafting of dissociated CNS tissue suspensions: a new approach for neuronal transplantation to deep brain sites. Brain Res 218:347–356
Dunnett SB, Björklund A (2000) Dissecting embryonic neural tissues for transplantation. In: Dunnett SB, Boulton AA, Baker GB (eds) Neural transplantation methods. Humana Press, Totowa, NJ, pp 3–25
Rosser AE, Barker RA, Armstrong RJE et al (2003) Staging and preparation of human fetal striatal tissue for neural transplantation in Huntington’s disease. Cell Transplant 12:679–686
Björklund A, Schmidt RH, Stenevi U (1980) Functional reinnervation of the neostriatum in the adult rat by use of intraparenchymal grafting of dissociated cell suspensions from the substantia nigra. Cell Tissue Res 212:39–45
Björklund A, Stenevi U, Schmidt RH et al (1983) Intracerebral grafting of neuronal cell suspensions. I. Introduction and general methods of preparation. Acta Physiol Scand Suppl 522:1–7
Kelly CM, Precious SV, Torres EM et al (2011) Medical terminations of pregnancy: a viable source of tissue for cell replacement therapy for neurodegenerative disorders. Cell Transplant 20:503–513
Precious SV, Zietlow R, Dunnett SB et al (2017) Is there a place for human fetal-derived stem cells for cell replacement therapy in Huntington’s disease? Neurochem Int 106:114–121
Barker RA, Mason SL, Harrower TP et al (2013) The long-term safety and efficacy of bilateral transplantation of human fetal striatal tissue in patients with mild to moderate Huntington’s disease. J Neurol Neurosurg Psychiatry 84:657–665
Polkinghorne J (1989) Review of the guidance on the research use of fetuses and fetal material. HM Stationery Office
Evtouchenko L, Studer L, Spenger C et al (1996) A mathematical model for the estimation of human embryonic and fetal age. Cell Transplant 5:453–464
Hurelbrink CB, Armstrong RJ, Barker RA et al (2000) Hibernated human fetal striatal tissue: successful transplantation in a rat model of Huntington’s disease. Cell Transplant 9:743–749
Hurelbrink CB, Tyers P, Armstrong RJE, Dunnett SB, Barker RA, Rosser AE (2003) Long-term hibernation of human fetal striatal tissue does not adversely affect its differentiation in vitro or graft survival: implications for clinical trials in Huntington’s disease. Cell Transplant 12:687–695
Piroth T, Pauly M-C, Schneider C et al (2014) Transplantation of human fetal tissue for neurodegenerative diseases: validation of a new protocol for microbiological analysis and bacterial decontamination. Cell Transplant 23:995–1007
Acknowledgments
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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Roberton, V.H., Rosser, A.E., McGorrian, AM., Precious, S.V. (2018). Dissection and Preparation of Human Primary Fetal Ganglionic Eminence Tissue for Research and Clinical Applications. In: Precious, S., Rosser, A., Dunnett, S. (eds) Huntington’s Disease. Methods in Molecular Biology, vol 1780. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7825-0_26
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DOI: https://doi.org/10.1007/978-1-4939-7825-0_26
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