Abstract
BackgroundBone Morphogenetic Protein (BMP) signaling pathways are involved in differentiation of stem cells into diverse cell types, and thus BMPs can be used as main guidance molecules for in vitro differentiation of human stem cells.Methodology/Principal FindingsWe have analyzed the ability for inducing differentiation of the heterodimer BMP-2/BMP-6 (BMP-2/6) compared to the homodimers BMP-2 or BMP-6, using human embryonic stem (hES) cells H9 as model system. When incubated in a medium with high concentration of basic fibroblastic growth factor (FGF2), 100 ng/ml of human recombinant BMPs induced morphological changes and differentiation of hES cells in 24 to 48 hours. After 5 days, expression of differentiation markers was induced and quantified by quantitative PCR (qPCR) and flow cytometry. BMP-2/6 exhibited stronger activity for the induction of the expression of trophectodermal (CDX2) and endodermal (SOX17, GATA4, AFP) markers than BMP-2 or BMP-6 homodimers. BMP-2/6 also induced the expression of BMPR2 gene more effectively than BMP-2 or BMP-6 when used at the same concentration and time. Moreover, the percentage of cells expressing the surface endodermal marker CXCR4 was also increased for the heterodimer when compared to both homodimers. BMP-2/6 was a more potent activator of Smad-dependent (SMAD1/5) and Smad-independent signaling (mitogen-activated protein kinases ERK and p38) than BMP-2 and BMP-6, and the activation of these pathways might play a role in its increased potency for inducing hES cell differentiation.Conclusions/SignificanceTherefore, we conclude that BMP-2/6 is more potent than BMP-2 or BMP-6 for inducing differentiation of hES cells, and it can be used as a more powerful substitute of these BMPs in in vitro differentiation guidance.
Highlights
Embryonic stem (ES) cell lines derived from the epiblast tissue of the inner cell mass of a blastocyst or earlier morula stage embryos are pluripotent and can develop to the three primary germ layers: ectoderm, endoderm and mesoderm
Forty-eight hours (h) after splitting, human embryonic stem (hES) cells were treated with Bone Morphogenetic Protein (BMP)-2, BMP-6 or BMP-2/6 at 100 ng/ml in mTeSR1 for 5 days, and the time course of morphological changes was analyzed
BMPs are known to be involved in several types of differentiation processes and the use of BMPs in differentiation of pluripotent cells is a powerful tool in biological and medical research [32,42]
Summary
Embryonic stem (ES) cell lines derived from the epiblast tissue of the inner cell mass of a blastocyst or earlier morula stage embryos are pluripotent and can develop to the three primary germ layers: ectoderm, endoderm and mesoderm. Murine and human stem cells display differences in their behavior upon incubation with BMPs. BMP signaling promotes self-renewal in mouse stem cells when incubated together with Leukemia Inhibitory Factor in the absence of serum replacement or conditioned medium [4]. Inhibition of BMP signaling is a requirement for long term maintenance of hES cells [5], while incubation with BMPs is a potent inductor of differentiation of these cells in conditions that would otherwise support self-renewal [6]. Bone Morphogenetic Protein (BMP) signaling pathways are involved in differentiation of stem cells into diverse cell types, and BMPs can be used as main guidance molecules for in vitro differentiation of human stem cells
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
