Late stage definitive endodermal differentiation can be defined by Daf1 expression

Abstract

BackgroundDefinitive endoderm (DE) gives rise to the respiratory apparatus and digestive tract. Sox17 and Cxcr4 are useful markers of the DE. Previously, we identified a novel DE marker, Decay accelerating factor 1(Daf1/CD55), by identifying DE specific genes from the expression profile of DE derived from mouse embryonic stem cells (ESCs) by microarray analysis, and in situ hybridization of early embryos. Daf1 is expressed in a subpopulation of E-cadherin + Cxcr4+ DE cells. The characteristics of the Daf1-expressing cells during DE differentiation has not been examined.ResultsIn this report, we utilized the ESC differentiation system to examine the characteristics of Daf1-expressing DE cells. We found that Daf1 expression could discriminate late DE from early DE. Early DE cells are Daf1-negative (DE-) and late DE cells are Daf1-positive (DE+). We also found that Daf1+ late DE cells show low proliferative and low cell matrix adhesive characteristics. Furthermore, the purified SOX17low early DE cells gave rise to Daf1+ Sox17high late DE cells.ConclusionDaf1-expressing late definitive endoderm proliferates slowly and show low adhesive capacity.Electronic supplementary materialThe online version of this article (doi:10.1186/s12861-016-0120-2) contains supplementary material, which is available to authorized users.

Defining Lineage-Specific Membrane Fluidity Signatures that Regulate Adhesion Kinetics

“…Adhesion strength of differentiated fibroblast has been known to be stronger than those of iPSCs ( Yu et al., 2018 , i.e., F iPSC < F MC ). In contrast, adhesion strength of late stage definitive endodermal cells showed gradual decreases according to the specific integrin subtypes ( Ogaki et al., 2016 , i.e., F HE < F iPSC ). Although both studies support the obtained order of adhesion strength, further studies are necessary to provide the conclusive answer.…”

Different murine-derived feeder cells alter the definitive endoderm differentiation of human induced pluripotent stem cells

“…The synthesized cDNA was used as a template for qPCR, which was performed using a SYBR Green real-time PCR Master Mix (TOYOBO, Osaka, Japan). The primers used for the undifferentiated cells, DE marker genes and hXIST RNA are shown in S1 Table [ 3 , 7 , 14 , 17 , 18 ]. PCR and data analyses were performed on an Applied Biosystems StepOne Plus real-time PCR system (Life Technologies).…”

“…In this study, we cultured the most widely used female hiPSC lines 201B7 and 253G1 [ 3 , 12 ] on SNL and MEF to evaluate their undifferentiated state and efficiency of differentiation into DE, depending on the type of feeder cell used. We found that the mRNA and protein expressions of the DE marker genes of the sex-determining region Y-box 17 (SOX17) and Forkhead box A2 (FOXA2) and the expressions of the DE surface marker C-X-C chemokine receptor type 4 (CXCR4) [ 7 , 13 , 14 ] were lower in the DE cells induced from 201B7 and 253G1 cells cultured on SNLs than in those induced from 201B7 and 253G1 cells cultured on MEFs under culture conditions employed for DE differentiation. These results suggested that undifferentiated culture of hiPSCs on SNLs inhibited DE differentiation when compared with MEFs.…”