ABSTRACT
The canonical Wnt/ß-catenin signaling pathway plays a crucial role in the maintenance of the balance between proliferation and differentiation throughout embryogenesis and tissue homeostasis. ß-Catenin, encoded by the Ctnnb1 gene, mediates an intracellular signaling cascade activated by Wnt. It also plays an important role in the maintenance of various types of stem cells including adult stem cells and cancer stem cells. However, it is unclear if ß-catenin is required for the derivation of mouse embryo-derived stem cells. Here, we established ß-catenin-deficient (ß-cat(Δ/Δ)) mouse embryo-derived stem cells and showed that ß-catenin is not essential for acquiring self-renewal potential in the derivation of mouse embryonic stem cells (ESCs). However, teratomas formed from embryo-derived ß-cat(Δ/Δ) ESCs were immature germ cell tumors without multilineage differentiated cell types. Re-expression of functional ß-catenin eliminated their neoplastic, transformed phenotype and restored pluripotency, thereby rescuing the mutant ESCs. Our findings demonstrate that ß-catenin has pleiotropic effects in ESCs; it is required for the differentiation of ESCs and prevents them from acquiring tumorigenic character. These results highlight ß-catenin as the gatekeeper in differentiation and tumorigenesis in ESCs.
Subject(s)
Carcinogenesis/genetics , Cell Differentiation/genetics , Embryonic Stem Cells/cytology , Embryonic Stem Cells/pathology , Genetic Pleiotropy , beta Catenin/metabolism , Animals , Blastocyst/cytology , Blastocyst/pathology , Embryonic Stem Cells/metabolism , Exons/genetics , Female , Gene Deletion , Male , Mice , Mice, Inbred C57BL , Teratoma/genetics , Teratoma/pathology , beta Catenin/deficiency , beta Catenin/geneticsABSTRACT
The capability of high-speed countercurrent chromatography (HSCCC) has been investigated for enrichment and determination of metal ions at trace levels. Separation of selected divalent metal ions was performed using a small coiled column. A hexane solution of 2-ethylhexylphosphonic acid mono-2-ethylhexyl ester (EHPA) was employed as the stationary phase. Loaded divalent metal ions such as Ni, Co, Cu, and Zn were chromatographically eluted in the order of increasing extractability by passing a mobile phase buffered at a desired pH. Individual metal ions showed good linearity between concentrations and chromatographic peak areas of the absorbance, as detected by postcolumn reaction with 4-(2-pyridylazo)resorcinol (PAR). Metal ions enriched into the stationary phase from a sample solution were separated into individual metal ions. The trace quantity of zinc in natural mineral water was determined by enrichment separation through an HSCCC column.
