[Optimization of RT-PCR conditions for estimation of peroxisomal genes [Catalase, PEX3] expression compare with pluripotent genes [Nanog, Oct4] expression in stem cells]

Peroxisomes are ubiquitous organelles in nearly all eukaryotes that participate in the metabolism of fatty acids and other metabolites. In order to investigate peroxisome biogenesis and gene profile expression analysis during neurogenesis, we have performed to set a semi-quantitative analysis for two peroxisomal genes expression [Catalase and PEX3] during neurogenesis comparing with stem cell marker genes [Oct4 and Nanog] in P19 cells. In this project we have used P19 cells which are suitable progenitor cells for the study of neurogenesis. Moreover, two different peroxisomal genes were selected to chase both kinetics of peroxisomal matrix protein synthesis [Catalase] and peroxisomal membrane protein biogenesis [Pex3p]. Total RNA extracted from P19 cells, and was used for cDNA synthesis at the next step. Specific primers of Oct4, Nanog, PEX3 and Catalase cDNAs were amplified by RT-PCR for quantitative analysis. Various parameters were changed to optimize the PCR profiles for each gene for further assessments. Here we are going to describe the conditions which we used, e.g.: PEX3 band was detected sharply after 35 cycles and at annealing temperature of 52.5-55.1°C. For PCR of Catalase we observed a sharp band after 35 cycles and annealing temperature of 52.7-55°C. Nanog cDNA was amplified after 36 cycles and between 60.1-64.1°C. Oct4 band was observed sharply after 28 cycles of amplification and annealing temperature in range of 53.4-57°C

[Review of stem cell usage in treatment of neurological diseases]

The stem cell possesses two distinct traits: self-renewal and differentiation capacity. The stem cell population is comprised of two main cell types: embryonic stem cells and adult stem cells. Researches on the stem cells provoked ways to utilize their potential for therapeutic treatments of multiple neurological disorders. There are three different approaches in neural repair with the usage of stem cells: cell replacements, neuroprotection and gene delivery which are highly developed in treating neurological disorders such as Parkinson and multiple sclerosis but these approaches have their own problems. In comparison with embryonic stem cells, adult stem cells have more application in neural repair, because the usage of embryonic stem cells raises ethical issues, and will involve allograft transplantation and are of much risk for tumor formation. Taking all of this into account, utilizing adult stem cells are in priority. Furthermore, injection of molecules such as bFGF and EGF leads to induction of neurogenesis that can be helpful in neural repair