RRC ID 67740
Author Joshi R, Tavana H.
Title Microengineered embryonic stem cells niche to induce neural differentiation.
Journal Annu Int Conf IEEE Eng Med Biol Soc
Abstract A major challenge in therapeutic use of embryonic stem cells (ESCs) for treating neurodegenerative diseases is creating a niche in vitro for controlled neural-specific differentiation of ESCs. We employ a niche microengineering approach to derive neural cells from ESCs by mimicking embryonic development in terms of direct intercellular interactions. Using a polymeric aqueous two-phase system (ATPS) microprinting technology, murine ESCs (mESCs) are precisely localized over a monolayer of supporting stromal cells to allow formation of individual mESC colonies. Polyethylene glycol (PEG) and dextran (DEX) are dissolved in culture media to form two immiscible aqueous solutions. A robotic liquid handler is used to print a nanoliter-volume drop of the denser DEX phase solution containing mESCs onto a confluent layer of supporting PA6 stromal cells submerged in the aqueous PEG phase. mESCs proliferate into isolated colonies of uniform size. For the first time, a comprehensive protein expression analysis of individual mESC colonies is performed over a two-week culture period to track temporal progression of cells from a pluripotent stage to specific neural cells. Starting from day 4, the expression of nestin, neural cell adhesion molecule (NCAM), and beta-III tubulin shows a significant increase but then levels off after the first week of culture. The expression of specific neural cell markers glial fibrillary acidic protein (GFAP), 2',3'-cyclic-nucleotide 3'-phosphodiesterase (CNPase), and tyrosine hydroxylase (TH) is elevated during the second week of culture. This microengineering approach to control ESCs differentiation niche combined with the time-course protein expression analysis of individual differentiating colonies facilitates understanding of evolution of specific neural cells from ESCs and identifying underlying molecular markers.
Volume 2015
Pages 3557-60
Published 2015-8-1
DOI 10.1109/EMBC.2015.7319161
PMID 26737061
MeSH 2',3'-Cyclic-Nucleotide Phosphodiesterases / metabolism Animals Biomarkers / metabolism Cell Differentiation Cells, Cultured Culture Media / chemistry Embryonic Stem Cells / cytology* Glial Fibrillary Acidic Protein / metabolism Mice Nestin / metabolism Neurons / cytology* Neurons / metabolism Stromal Cells Tyrosine 3-Monooxygenase / metabolism
Human and Animal Cells MC3T3-G2/PA6(RCB1127)