Stem Cells and Parkinson's Disease

comparative Essay
2615 words
2615 words

The goal of this paper is to compare the utility of adult, embryonic and induced pluripotent stem cells (iPSCs) to treat Parkinson’s disease. As such several things will be assessed, dosage of stemcells, improvement in motor function, in combination with the presence of α-synuclein proteins and cell survival. To give a short overview of the steps that will be taken to complete the study. Obtaining stem cells, whether adult, embryonic or induced, shall be done using healthy mouse models and after ethical approval has been gained. The process to derive them will be detailed below, however they are also purchasable commercially with the benefit of being well studied and accompanied by a detailed analysis of properties, however with a higher purchasing costs. All steps should be done using sterile techniques to avoid contamination, both to object and self. All (disposable) materials should be sterilized and (where possible) only for 1 time use, due to the sensitivity stem cells have to detergents. After verifying they are bonafide stem cells the next phase should be ready to commence. Rats will be used as a disease model. All will be healthy, adult and a baseline PET scan and performance on a rotarod will be done. All but 1 set will then be subjected to a neurotoxin to mimic the diseased Parkinson state. Another PET scan and rotarod test will be done on all groups. The groups affected by the neurotoxin will then be subdivided into those receiving the different types of stem cells, one subgroup will receive no treatment and one group will receive medium (no cells). Next to this 2 different dosages will occur. Another PET scan and rotarod will be done to determine if any improvement occurs. Rats will then be sacrificed to determi... ... middle of paper ... ...ter a 6-OHDA lesion of the substantia nigra in a single hemisphere and if the injected cells would become infected by α-synuclein proteins. References: Bjorklund LM, Sánchez-Pernaute R, Chung S, Andersson T, Chen IY, McNaught KS, Brownell AL, Jenkins BG, Wahlestedt C, Kim KS, et al. Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model. Proc Natl Acad Sci USA. 2002;99:2344–2349. Lerou, P.H. et al. Derivation and maintenance of human embryonic stem cells from poor-quality in vitro fertilization embryos. Nat. Protoc. 3, 923– 933 (2008). Mead R. 1988. The design of experiments. Cambridge, New York: Cambridge University Press. 620 p. Takahashi, K., Okita, K., Nakagawa, M. & Yamanaka, S. Induction of pluripotent stem cells from fibroblast cultures. Nat. Protoc. 2, 3081–3089 (2007).

In this essay, the author

  • Recommends using mef medium with fbs instead of sr-medium to disaggregate the embryo.
  • Explains that ipscs can be obtained from several different cell types. adipose cells are a viable alternative to fibroblasts with high accessibility in pd patients.
  • Explains how to obtain adipose cells by removing fat pads from 8-10 week old mice and placing them in hanks balanced salt solution.
  • Explains that the use of neurotoxin rodent models to study parkinson's has been well established. 6-ohda causes oxidative stress to daergic neurons, causing them to die quickly.
  • Recommends a post-hoc analysis to determine if tukey’s hsd test would give the result that most benefitted from the admission of stem cells.
  • Explains that different types of stem cells are being investigated in animal models in order to find a cure for parkinson’s disease.
  • Outlines the steps involved in the study to compare the utility of adult, embryonic and induced pluripotent stem cells (ipscs) to treat parkinson's disease.
  • Explains the culturing of mouse and human embryonic stem cells using dmem, l-glutamine, penicillin/streptomycin, and knockout sr-medium.
  • Explains how to produce the retrovirus using 293t cells, dmem, and pmx, gag-pol and vsv-g–expressing plasmids.
  • Explains that the ventricles provide the most neurospheres, but the subventricular zone is of particular importance since it may represent a source in aged and parkinson’s affected individuals.
  • Explains that mrna transcripts can be used to determine cell differentiation status, and how far differentiated a mixture of cells may be.
  • Explains that rats will be injected with immunosuppressors to prevent rejection. a lower dose of embryonic stem cells 1,000-2,000 cells / l might be more beneficial due to more direct contact with the neuronal environment.
  • Explains that several moments of rotational behavior assessment will need to take place across several different groups to assess the effects of the stem cells.
  • Recommends using mead's resource equation to estimate the group size. the statistical test would be a two-way-anova to analyse the rotarod performance.
  • Explains that embryonic stem cells develop into functional dopaminergic neurons after transplantation in a parkinson rat model.
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