RRC ID 65744
Author Adnan N, Mie M, Haque A, Hossain S, Mashimo Y, Akaike T, Kobatake E.
Title Construction of a Defined Biomimetic Matrix for Long-Term Maintenance of Mouse Induced Pluripotent Stem Cells.
Journal Bioconjug Chem
Abstract The existing in vitro culture systems often use undefined and animal-derived components for the culture of pluripotent stem cells. Artificial bioengineered peptides have the potential to become alternatives to these components of extracellular matrix (ECM). Integrins and cadherins are two cell adhesion proteins important for stem cell self-renewal, differentiation, and phenotype stability. In the present study, we sought to mimic the physico-biochemical properties of natural ECMs that allow self-renewal of mouse induced pluripotent stem cells (iPSCs). We develop a genetically engineered ECM protein (ERE-CBP) that contains (i) an integrin binding peptide sequence (RGD/R), (ii) an E-/N-cadherin binding peptide sequence (SWELYYPLRANL/CBP), and (iii) 12 repeats of APGVGV elastin-like polypeptides (ELPs/E).While ELPs allow efficient coating by binding to nontreated hydrophobic tissue culture plates, RGD/R and CBP support integrin- and cadherin-dependent cell attachment, respectively. Mouse iPSCs on this composite matrix exhibit a more compact phenotype compared to cells on control gelatin substrate. We also demonstrated that the ERE-CBP supports proliferation and long-term self-renewal of mouse iPSCs for up to 17 passages without GSK3β (CHIR99021) and Erk (PD0325901) inhibitors. Overall, our engineered ECM protein, which is cost-effective to produce in prokaryotic origin and flexible to modify with other cell adhesion peptides or growth factors, provides a novel approach for expansion of mouse iPSCs in vitro.
Volume 27(7)
Pages 1599-605
Published 2016-7-20
DOI 10.1021/acs.bioconjchem.6b00141
PMID 27269811
MeSH Adsorption Amino Acid Sequence Animals Biomimetics / methods* Cell Culture Techniques / methods* Cell Differentiation / drug effects Cell Self Renewal / drug effects Extracellular Matrix Proteins / chemistry Extracellular Matrix Proteins / genetics Extracellular Matrix Proteins / metabolism Extracellular Matrix Proteins / pharmacology Hydrophobic and Hydrophilic Interactions Induced Pluripotent Stem Cells / cytology* Induced Pluripotent Stem Cells / drug effects* Integrins / metabolism Mice Protein Engineering
IF 4.031
Human and Animal Cells iPS-MEF-Ng-178B-5(APS0002)