Ps-50: Derivation of Definitive Endoderm from hiPSCs Cultured on PLA Nanofibrous Scaffolds

Hoveizi E *, Nabiuni M , Massumi M , Parivar K , Ai J ,


Objective: Over the last two decades, whether cell replacement therapy can overcome the using of insulin administration in diabetes mellitus type 1 (IDDM) patients or not, has became a challenging issue amongst clinicians and cell biologists. The idea to generate human induced pluripotent stem cells (hiPSCs) specific from diabetic patient and differentiated to insulin-producing cells pioneered by Melton’s lab, has opened a new window to using of iPS cells as new source for cell therapy of diabetes without warring about the immunological rejection or ethical issues. Use of 3 dimensional (3D) scaffolds is one access to potentially enhance common differentiation protocols, which has been shown to improve cellular function and differentiation potential. Materials and Methods: In the present study we intended to using pluripotent hiPS seeded onto Matrigel coated 3D poly lactic acid (PLA) scaffolds to derive definitive endoderm cells for 7 days, the first crucial stage of endoderm-derived tissues differentiation by use of Activin A with concentration of 100 ng/ml and Wnt3a with concentration of 25 ng/ml. We used immunofluorescent and RT-PCR methods for survey experiments. Results: Our results showed that hiPS which were coaxed to differentiate on Matrigel coated 3D scaffolds can be in prosperous manner enticed to differentiate into definitive endoderm. The immunofluorescent study and RT-PCR results revealed that iPS cells that were cultured on Matrigel coated 3D scaffolds expressed significantly higher levels of the key endoderm transcription factors SOX17 and GSC in comparison to those differentiated on 2D cultures. Conclusion: Our research confirmed the positive effect of 3D cultures on endoderm commitment of hiPS. The result of this study may have impact in future therapy of IDDM patien by cell replacement therapy.