Direct Conversion of Human Umbilical Vein Endothelial Cells into Osteoblasts Using Cell Penetrating Peptide, 30Kc19
Mammalian Synthetic Biology Workshop
2018
Fifth International Mammalian Synthetic Biology Workshop
Poster Session
Poster Session
Saturday, May 5, 2018 - 5:30pm to 6:30pm
In this study, osteogenic transcription factors, Osterix (Osx), and Core-Binding Factor Beta (Cbfb), are individually attached to a CPP, 30Kc19, to transdifferentiate HUVEC into osteoblasts. Osx and Cbfb are key ostegenic transcription factors known for their roles in osteoblast maturation, bone calcification, and bone homeostasis. In order to attach 30Kc19 to these transcription factors, gene cloning via recombinant technology and mass production using BL21 Escherichia coli are employed. Further promoting transdifferentation, Bone Morphogenic Protein 4 (BMP4) is incorporated to induce osteogenesis from different cell types, such as HUVEC. Prior to the peptide treatments, direct reprogramming efficacies in HUVEC and Adipocyte Derived Stem Cell (ADSC) are examined via electroporation of Osx plasmid. When transfected HUVEC and ADSC are analyzed, calcification and up-regulation of osteoblast-specific genes are observed, indicating that transdifferentiation occurred.