Gustavo Jesús Vázquez Zapién
Instituto Politécnico Nacional, México
Title: FTIR spectroscopic and molecular analysis of differentiated pancreatic cells from pluripotent stem cells
Biography
Biography: Gustavo Jesús Vázquez Zapién
Abstract
An essential property of stem cells (SCs) is the potentiality, which refers to their capability to differentiate into different cell lineages under certain culture conditions. Two of the greatest challenges in SCs biology and regenerative medicine are differentiation control of SCs and to ensure the purity of differentiated cells. In this work, mouse Pluripotent Stem Cells (mPSCs) were subjected to a differentiation protocol of 21 days obtaining Differentiated Pancreatic Cells (DPCs), characterizing this differentiation process by spectroscopic and genetic analysis through Fourier Transform Infrared (FTIR) spectroscopy and Real-Time PCR. FTIR spectra of mPSCs and DPCs at different maturation stages (11, 17 and 21 days) were obtained and showed absorption bands related with different types of biomolecules. These FTIR spectra exhibited significant changes particularly in the proteins, carbohydrates and nucleic acids bands agreeing with the differentiation process. The second derivative of these bands and the Principal Components Analysis allowed us to compare and discriminate mPSCs from DPCs. About gene expression, mPSCs expressed pluripotency genes (Nanog and SOX2) and the DPCs expressed endodermal genes (SOX17 and Pdx1) at day 11, an inductor gene of embryonic pancreas development (Pdx1) at day 17 and pancreas genes (Insulin and Glucagon) at day 21 of differentiation. In conclusion, we obtained DPCs from mPSCs, which passed through the chronological stages of embryonic pancreas development. The FTIR spectral changes provide a new biophysical parameter based on molecular markers that indicate the differentiation process of mPSCs to specialized cells.