الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
 |  | from | 86 |  |  |
| | | from | 86 | | |
Abstract
Annually, neurodegenerative disease mortality increases by 119-231% in the USA in the period 1990–2040 (282). To date there is no definitive treatment for neural loss in neurodegenerative diseases. Available treatment is considered as symptomatic treatment; therefore, it is important to find other alternatives such as stem cell-based neural regeneration therapy.
Stem cell-based neural regeneration therapy has recently gained extensive attention, being one of the most promising treatments for the damaged neural tissue. Stem cells are a class of undifferentiated cells capable of self-renewal and multilineage differentiation. This occurs through a process of asymmetric mitosis that leads to two daughter cells, one identical to the stem cell and one capable of differentiation into a more mature cell when given the appropriate environmental cues.
Among the well-known differentiation molecules, TH and Curcumin are important small molecules that could stimulate differentiation of non-neural precursor cells into neural cells via up-regulation of neural lineage confined TFs.
The present work aimed to standardize a protocol ensuring the best differentiation of bone marrow stromal cells (HS5) into functional neural cells by exposure of stem cells to BME, TH, and curcumin.
This work was performed on HS5 cells which were purchased bone marrow stromal cells from stem cell technology. This study was done at Pharmaceutical Research institute (PRI), Albany College of pharmacy and Health science, New York, USA in time period from 15th March to 15th July 2019.
Proliferated HS5 cells received pre-induction media composed of CCM with 20% FBS/ 1 mM BME. 24 hour later, cells were transferred to neuronal induction media composed of DMEM and 1 mM BME, T3 0,5 µM, curcumin 5 µM. Then cells were assessed at 3 time points, 24h, 48 h, and 7 days for PAX6 assessment with flow cytometry. 7 days post neural induction PAX6, SOX2, DLX2, and, GAP-43 were assessed with flow cytometry and immunofluorescence in addition to nissl staining of differentiated cells. Nuclear immunostaining with PI was assessed in differentiated cells 7 days post neural induction of HS5 cells with BME, T3, and curcumin. Another group of HS5 cells were induced for neural differentiation with BME, T3, and curcumin without BME pre-conditioning. For this group PAX6 was assessed b flowcytometry 48 h post neural induction.
The results showed that PAX6 expression is increased significantly with BME, T3, and Curcumin at 48h time point in comparison to the 24 h time point. In addition, PAX6 was significantly suppressed with these molecules at the 7-day. Along with PAX6, DLX2 was significantly expressed with BME, T3, and curcumin 7 days post neural induction
Our results revealed at 7 days post neural induction positive expression of SOX2 with BME and curcumin while T3 suppress its levels, in accordance with SOX2, BME and Curcumin revealed positive expression of GAP-43 while T3 showed no effect
Nuclear staining was assessed by PI staining showing evident nuclear with curcumin and to a lesser extent with T3 while BME had bad impacts on cell viability.
In addition, our results showed differentiated neural cells having nissl bodies in their cell bodies under the effect of T3 and curcumin while BME didn’t have a clear effect on nissl bodies staining.