Pancreatic Transcription Factors Containing Protein Transduction Domains Drive Mouse Embryonic Stem Cells towards Endocrine Pancreas

• DOI: 10.1371/journal.pone.0036481
• Publication Date: 2012-05-01T20:58:17Z
• Authors: Maria João Lima; Hilary M. Docherty; Yuanxiao Chen; Ludovic Vallier; Kevin Docherty
• Keywords: PDX1; PAX4; Embryoid body

Protein transduction domains (PTDs), such as the HIV1-TAT peptide, have been previously used to promote uptake of proteins into a range cell types, including stem cells. Here we generated pancreatic transcription factors containing PTD sequences and administered these endoderm enriched mouse embryonic (ES) cells under conditions that were designed mimic pattern expression in developing pancreas. The ES first cultured embryoid bodies treated with Activin A Bone morphogenetic protein 4 (BMP4) formation definitive endoderm. Cells subsequently plated monolayer different combinations modified recombinant Pdx1 MafA. results demonstrate each factor was efficiently taken up by cells, where they localized nuclei. RT-qPCR measure levels markers. After addition alone for period five days, followed combination TAT-MafA second phase, up-regulation insulin 1, 2, Pdx1, Glut2, Pax4 Nkx6.1 observed. As assessed immunocytochemistry, double positive detected differentiated cultures. Although markers cultures comparable transformed β-cell line (MIN-6) human islets, observed several orders magnitude lower. This suggests that, although PTD-TFs may prove useful studying role exogenous TFs differentiation towards islets other lineages, amount is well below required therapeutically