Ps-75: Generation and Maintenace of Human Embryonic Stem Cells-Derived Neural Precursors Cells by Small Molecules


Nemati Sh *, Karamali F , Kiani S , Baharvand H ,

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Objective: Human embryonic stem cells (hESCs) have led to an important revolution in stem cell research and regenerative medicine. In order to create specific neural progenitors (NPs), we have established a homogenous, expandable and self-renewable population of multipotent NPs from hESCs using an adherent system and defined medium supplemented with a combination of small molecules. Materials and Methods: Here, after generation of NPs, we characterized them and evaluated their potency by Real time PCR, Immuno fluorecense and Flow cytometery satining, karyotype and pacth clamp analysis. Results: The established hESCs-NPs highly expressed Nestin and Sox1 and Pax6. These NPs were continuously propagated for approximately six months without losing their potential to generate astrocytes, oligodendrocytes, and functional neurons, and maintained a stable chromosome number. Voltage clamp analysis revealed outward potassium currents in hESC-NPs. The self-renewal characteristic of the NPs was demonstrated by a symmetrical mode of Nestin-positive cell division. Additionally, these hiPSC-NPs can be easily frozen and thawed in the presence of ROCK inhibitor without losing their proliferation, karyotype stability, and developmental potential. Conclusion: The characteristics of our generated hESC-NPs provide the opportunity to use patient-specific or ready-to-use hESC-NPs in future biomedical applications.