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Stem Cells International
Volume 2016, Article ID 6709714, 10 pages
http://dx.doi.org/10.1155/2016/6709714
Research Article

FTIR Spectroscopic and Molecular Analysis during Differentiation of Pluripotent Stem Cells to Pancreatic Cells

1CIBA-Tlaxcala, Instituto Politécnico Nacional, 90700 Tepetitla, TLAX, Mexico
2Laboratorio de Embriología, Escuela Médico Militar, Universidad del Ejército y Fuerza Aérea, 11200 Ciudad de México, Mexico
3Laboratorio de Biología Celular y Tisular, Escuela Médico Militar, Universidad del Ejército y Fuerza Aérea, 11200 Ciudad de México, Mexico
4Laboratorio Multidisciplinario de Investigación, Escuela Militar de Graduados de Sanidad, Universidad del Ejército y Fuerza Aérea, 11200 Ciudad de México, Mexico
5Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional, 07320 Ciudad de México, Mexico

Received 9 March 2016; Revised 12 July 2016; Accepted 20 July 2016

Academic Editor: Leonard M. Eisenberg

Copyright © 2016 Gustavo Jesus Vazquez-Zapien et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Some of the greatest challenges in stem cells (SCs) biology and regenerative medicine are differentiation control of SCs and ensuring the purity of differentiated cells. In this work, we differentiated mouse pluripotent stem cells (mPSCs) toward pancreatic cells characterizing this differentiation process by molecular and spectroscopic technics. Both mPSCs and Differentiated Pancreatic Cells (DPCs) were subjected to a genetic, phenotypic, and biochemical analysis by real-time quantitative PCR (RT-qPCR), immunocytochemistry, and Fourier Transform Infrared (FTIR) spectroscopy. Cultured mPCSs expressed pluripotent genes and proteins (Nanog and SOX2). DPCs expressed endodermal genes (SOX17 and Pdx1) at day 11, an inductor gene of embryonic pancreas development (Pdx1) at day 17 and pancreas genes and proteins (Insulin and Glucagon) at day 21 of differentiation. Likewise, 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 spectral changes agreeing with the differentiation process, particularly in proteins and nucleic acids bands. In conclusion, the obtained DPCs passed through the chronological stages of embryonic pancreas development and FTIR spectra provide a new biophysical parameter based on molecular markers indicating the differentiation process of mPSCs to specialized cells.