Conversion of human fibroblasts into neuro sphere-like cells using a single polycistronic vector
9th Biotechnology Congress
August 31-September 02, 2015 Orlando,Florida, USA

Fahimeh Mirakhori and Hossein Baharvand

Royan Institute for Stem Cell Biology and Technology, Iran

Posters-Accepted Abstracts: J Biotechnol Biomater

Abstract:

The unique capability of neural progenitor cells to induce regeneration in several animal models of neurological disorders, make them the best potential cell source for regenerative medicine. To date, direct lineage conversion has shown many distinct advantages in the field of cellular reprogramming to capture the patient specific neuronal progenitor cells on a dish to enhance the knowledge of basic processes of human disease, development and drug discovery. A number of studies have demonstrated that transient expression of Yamanaka factors with appropriate supporting signals can efficiently convert fibroblasts into NPCs�??called iPSC-factor�??based reprogramming. Here, we used a minimalist approach combining a single polycistronic doxycycline (dox)-inducible lentiviral vector containing SOX2, OCT4, cMYC and KLF4 with small molecule inhibitors to convert human fibroblasts into NPC-like cells. We limited the exogenous expression of Yamanaka factors for 14 days. The treated human fibroblast cells exhibit neurosphere- like morphology within 28 days post transduction. The immunostaining of the resulted cells showed that they were positive for neural markers: Sox2, PAX6 and NESTIN. However, under the same defined condition, no colonies formed from transduced human fibroblasts when weexpunged these small molecules from our induction regime. This finding showed the important power of developmental signals which could enable partially converted human fibroblast cells morphed into neural progenitor-like cells. The present study is the first demonstration that the presence of neural inducer chemical dual SMAD inhibitors can mediate human fibroblasts reprogramming to NPClike cells via a single polycistronic vector of Yamanaka factors.

Biography :

Email: mirakhori@ut.ac.ir