EDA-Containing Fibronectin Increases Proliferation of Embryonic Stem Cells

Abstract

Embryonic stem cells (ESC) need a set of specific factors to be propagated. They can also grow in conditioned medium (CM) derived from a bovine granulosa cell line BGC (BGC-CM), a medium that not only preserves their main features but also increases ESC´s proliferation rate. The mitogenic properties of this medium were previously reported, ascribing this effect to an alternative spliced generated fibronectin isoform that contains the extra domain A (FN EDA+). Here, we investigated if the FN EDA+ isoform increased proliferation of mouse and human ES cells. We analyzed cell proliferation using conditioned media produced by different mouse embryonic fibroblast (MEF) lines genetically engineered to express FN constitutively including or excluding the EDA domain (FN EDA-), and in media supplemented with recombinant peptides containing or not the EDA. We found that the presence of EDA in the medium increased mouse and human ESC’s proliferation rate. Here we showed for the first time that this FN isoform enhances ESC’s proliferation. These findings suggest a possible conserved behavior for regulation of ES cells proliferation by this FN isoform and could contribute to improve their culturing conditions both for research and cell therapy.

EDA fibronectin–TLR4 axis sustains megakaryocyte expansion and inflammation in bone marrow fibrosis

“…Anti–phospho-ERK 1/2, anti-ERK 1/2, anti–phospho–STAT-5, anti–STAT-5, anti–phospho p65 (NF-κB), and p65 (NF-κB) were purchased from Cell Signaling Technologies. EDA + and EDA − recombinant peptides were produced as previously described (Losino et al, 2013). All the experiments were performed with the same batch for each peptide.…”

Manganese Superoxide Dismutase Gene Expression Is Induced by Nanog and Oct4, Essential Pluripotent Stem Cells’ Transcription Factors

“…R1 and Ainv15 ESC lines are commercial lines purchased from ATCC. They were cultured and differentiated as previously described [ 13 – 15 ]. The iPSC-20 line was derived and validated previously by us [ 16 ], and cultured and differentiated as previously described [ 16 ].…”

Thermo‐modulated Hela cell release from an elastic and biocompatible hydrogel

“…This P(AEtMA‐Cl‐DEAEA)/PA scaffolds responded slowly to the ambient temperature. The diad hydrogels became much more swollen at lower temperatures (10.6× at 20°C and 8.6× at 37°C in PBS) and the changes in physical propertied of the polymers may trigger the binding proteins to release at low temperatures . Cells were unable to find peptide ligands and consequently detached.…”