Abstract
Pluripotent stem cells convert into skeletal muscle tissue during teratoma formation or chimeric animal development. Thus, they are characterized by naive myogenic potential. Numerous attempts have been made to develop protocols enabling efficient and safe conversion of pluripotent stem cells into functional myogenic cells in vitro. Despite significant progress in the field, generation of myogenic cells from pluripotent stem cells is still challenging—i.e., currently available methods require genetic modifications, animal-derived reagents, or are long lasting—and, therefore, should be further improved. In the current study, we investigated the influence of interleukin 4, a factor regulating inter alia migration and fusion of myogenic cells and necessary for proper skeletal muscle development and maintenance, on pluripotent stem cells. We assessed the impact of interleukin 4 on proliferation, selected gene expression, and ability to fuse in case of both undifferentiated and differentiating mouse embryonic stem cells. Our results revealed that interleukin 4 slightly improves fusion of pluripotent stem cells with myoblasts leading to the formation of hybrid myotubes. Moreover, it increases the level of early myogenic genes such as Mesogenin1, Pax3, and Pax7 in differentiating embryonic stem cells. Thus, interleukin 4 moderately enhances competence of mouse pluripotent stem cells for myogenic conversion.
Highlights
Pluripotent stem cells (PSCs), such as embryonic stem cells (ESCs) or induced pluripotent stem cells, are characterized by unique properties, i.e., the ability of extensive proliferation, self-renewal, as well as multidirectional differentiation enabling generation of all mammalian cell types
Activation of canonical Wnt, followed by fibroblast growth factor 2 (FGF-2) treatment led to the derivation of myogenic cells with high efficiency, i.e., nearly 90% of cells derived from mouse or human ESCs synthesized myogenic markers—such as Pax3, the marker of skeletal muscle precursor cells formed during embryonic myogenesis, or myosin heavy chains (MHC)—characteristic of already committed and differentiated myogenic cells [5]
We showed that undifferentiated ESCs lack proteins important for skeletal muscle myoblast fusion such as vascular cell adhesion molecule 1 (VCAM1) and M-cadherin, which may limit their competence for fusion [18]
Summary
Pluripotent stem cells (PSCs), such as embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs), are characterized by unique properties, i.e., the ability of extensive proliferation, self-renewal, as well as multidirectional differentiation enabling generation of all mammalian cell types. On the basis of previously described data on IL-4’s role in myogenic and other cells, we investigated the influence of this cytokine on PSCs to verify whether IL-4 may enhance ESC ability to fuse with myoblasts as well as other features of these cells, e.g., proliferation rate or their gene expression profile. We analyzed both undifferentiated ESCs and cells cultured in embryoid bodies (EBs), i.e., three dimensional aggregates formed by PSCs differentiating in suspension culture, in medium lacking self-renewal promoting factors, such as leukemia inhibitory factor (LIF)
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