Generation and Characterization of Induced Pluripotent Stem Cells Derived from Parkinson's Disease Patients with Familial SNCA A53T Mutation and Sporadic Cases

Abstract

Nowadays, induced pluripotent stem cells (iPSCs) are the most highlighted type of stem cells in research as they overcome the limitations associated with embryonic stem cells (ESCs) usage and hold a promising future for disease modeling and treatment. Generation of iPSCs is dependent on reprogramming of somatic cells by the forced introduction of reprogramming factors into the cell. To date, reprogramming procedure remains inefficient process, since a small portion of transfected cells can undergo complete reprogramming. One of the identified factors that affect the reprogramming efficiency is donor age. In this thesis, we aim to test the effect of donor age on reprogramming process using skin fibroblasts collected from PD patients who were diagnosed with early onset familial PD (A53T mutation) and late onset sporadic PD. To achieve this goal, fibroblasts were reprogrammed using Sendai virus vector to generate iPSCs. All the generated iPSCs were compared to ESCs and the commercially available iPSCs to ensure that these generated iPSCs have the same characteristic features and to confirm their ability to express pluripotent protein markers. The number of emerged colonies was used to reflect the reprogramming efficiency. In our study, we observed that A53T (2) sample showed the highest number of iPSC colonies among the other reprogrammed samples. Our results showed that the donor age had no effect on our reprogramming procedure as number of emerged iPSC colonies in sporadic PD (age 69) was similar to both control and other A53T samples (age 45 and 48, respectively) that were isolated from individuals who were similar in age with A53T (2) sample. Therefore, further transcriptomic analysis is required to identify intrinsic factor(s) that may play a role in the significantly high number of iPSC colonies observed with A53T (2) reprogramming.

Keywords Stem cells, Parkinson, Donor age, Intrinsic factors