Derivation of Myogenic Progenitors Directly From Human Pluripotent Stem Cells Using a Sphere-Based Culture

Stem cells translational medicine - Tập 3 Số 5 - Trang 564-574 - 2014
Tohru Hosoyama1, Jered V. McGivern2, Jonathan M. Van Dyke1, Allison D. Ebert2, Masatoshi Suzuki1
1Department of Comparative Biosciences and The Stem Cell and Regenerative Medicine Center, University of Wisconsin, Madison, Wisconsin, USA
2Department of Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, USA

Tóm tắt

Abstract Using stem cells to replace degenerating muscle cells and restore lost skeletal muscle function is an attractive therapeutic strategy for treating neuromuscular diseases. Myogenic progenitors are a valuable cell type for cell-based therapy and also provide a platform for studying normal muscle development and disease mechanisms in vitro. Human pluripotent stem cells represent a valuable source of tissue for generating myogenic progenitors. Here, we present a novel protocol for deriving myogenic progenitors from human embryonic stem (hES) and induced pluripotent stem (iPS) cells using free-floating spherical culture (EZ spheres) in a defined culture medium. hES cell colonies and human iPS cell colonies were expanded in medium supplemented with high concentrations (100 ng/ml) of fibroblast growth factor-2 (FGF-2) and epidermal growth factor in which they formed EZ spheres and were passaged using a mechanical chopping method. We found myogenic progenitors in the spheres after 6 weeks of culture and multinucleated myotubes following sphere dissociation and 2 weeks of terminal differentiation. A high concentration of FGF-2 plays a critical role for myogenic differentiation and is necessary for generating myogenic progenitors from pluripotent cells cultured as EZ spheres. Importantly, EZ sphere culture produced myogenic progenitors from human iPS cells generated from both healthy donors and patients with neuromuscular disorders (including Becker's muscular dystrophy, spinal muscular atrophy, and familial amyotrophic lateral sclerosis). Taken together, this study demonstrates a simple method for generating myogenic cells from pluripotent sources under defined conditions for potential use in disease modeling or cell-based therapies targeting skeletal muscle.

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Tài liệu tham khảo

Hosoyama, 2012, Applications of skeletal muscle progenitor cells for neuromuscular diseases, Am J Stem Cells, 1, 253

Tedesco, 2012, Stem cell therapies for muscle disorders, Curr Opin Neurol, 25, 597, 10.1097/WCO.0b013e328357f288

Salani, 2012, Generation of skeletal muscle cells from embryonic and induced pluripotent stem cells as an in vitro model and for therapy of muscular dystrophies, J Cell Mol Med, 16, 1353, 10.1111/j.1582-4934.2011.01498.x

Rao, 2012, Highly efficient derivation of skeletal myotubes from human embryonic stem cells, Stem Cell Rev, 8, 1109, 10.1007/s12015-012-9413-4

Barberi, 2007, Derivation of engraftable skeletal myoblasts from human embryonic stem cells, Nat Med, 13, 642, 10.1038/nm1533

Awaya, 2012, Selective development of myogenic mesenchymal cells from human embryonic and induced pluripotent stem cells, PLoS One, 7, e51638, 10.1371/journal.pone.0051638

Darabi, 2008, Functional skeletal muscle regeneration from differentiating embryonic stem cells, Nat Med, 14, 134, 10.1038/nm1705

Darabi, 2011, Assessment of the myogenic stem cell compartment following transplantation of Pax3/Pax7-induced embryonic stem cell-derived progenitors, Stem Cells, 29, 777, 10.1002/stem.625

Darabi, 2012, Human ES- and iPS-derived myogenic progenitors restore DYSTROPHIN and improve contractility upon transplantation in dystrophic mice, Cell Stem Cell, 10, 610, 10.1016/j.stem.2012.02.015

HD iPSC Consortium, 2012, Induced pluripotent stem cells from patients with Huntington's disease show CAG-repeat-expansion-associated phenotypes, Cell Stem Cell, 11, 264, 10.1016/j.stem.2012.04.027

Ebert, 2009, Induced pluripotent stem cells from a spinal muscular atrophy patient, Nature, 457, 277, 10.1038/nature07677

Ebert, 2013, EZ spheres: A stable and expandable culture system for the generation of pre-rosette multipotent stem cells from human ESCs and iPSCs, Stem Cell Res (Amst), 10, 417, 10.1016/j.scr.2013.01.009

Sareen, 2012, Inhibition of apoptosis blocks human motor neuron cell death in a stem cell model of spinal muscular atrophy, PLoS One, 7, e39113, 10.1371/journal.pone.0039113

Tamaki, 2003, Growth and differentiation potential of main- and side-population cells derived from murine skeletal muscle, Exp Cell Res, 291, 83, 10.1016/S0014-4827(03)00376-8

Tamaki, 2008, Skeletal muscle-derived CD34+/45− and CD34−/45− stem cells are situated hierarchically upstream of Pax7+ cells, Stem Cells Dev, 17, 653, 10.1089/scd.2008.0070

Hosoyama, 2013, Isolation and in vitro propagation of human skeletal muscle progenitor cells from fetal muscle, Cell Biol Int, 37, 191, 10.1002/cbin.10026

Sarig, 2006, Regeneration and transdifferentiation potential of muscle-derived stem cells propagated as myospheres, Stem Cells, 24, 1769, 10.1634/stemcells.2005-0547

Wei, 2011, Human skeletal muscle-derived stem cells retain stem cell properties after expansion in myosphere culture, Exp Cell Res, 317, 1016, 10.1016/j.yexcr.2011.01.019

Westerman, 2010, Adult muscle “stem” cells can be sustained in culture as free-floating myospheres, Exp Cell Res, 316, 1966, 10.1016/j.yexcr.2010.03.022

Park, 2008, Disease-specific induced pluripotent stem cells, Cell, 134, 877, 10.1016/j.cell.2008.07.041

Mitne-Neto, 2011, Downregulation of VAPB expression in motor neurons derived from induced pluripotent stem cells of ALS8 patients, Hum Mol Genet, 20, 3642, 10.1093/hmg/ddr284

Yu, 2007, Induced pluripotent stem cell lines derived from human somatic cells, Science, 318, 1917, 10.1126/science.1151526

Thomson, 1998, Embryonic stem cell lines derived from human blastocysts, Science, 282, 1145, 10.1126/science.282.5391.1145

Ludwig, 2006, Feeder-independent culture of human embryonic stem cells, Nat Methods, 3, 637, 10.1038/nmeth902

Ludwig, 2006, Derivation of human embryonic stem cells in defined conditions, Nat Biotechnol, 24, 185, 10.1038/nbt1177

Svendsen, 1998, A new method for the rapid and long term growth of human neural precursor cells, J Neurosci Methods, 85, 141, 10.1016/S0165-0270(98)00126-5

Nelson, 2008, A high concentration of epidermal growth factor increases the growth and survival of neurogenic radial glial cells within human neurosphere cultures, Stem Cells, 26, 348, 10.1634/stemcells.2007-0299

Nelson, 2006, Low concentrations of extracellular FGF-2 are sufficient but not essential for neurogenesis from human neural progenitor cells, Mol Cell Neurosci, 33, 29, 10.1016/j.mcn.2006.06.003

Takahashi, 2006, Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors, Cell, 126, 663, 10.1016/j.cell.2006.07.024

Takahashi, 2007, Induction of pluripotent stem cells from adult human fibroblasts by defined factors, Cell, 131, 861, 10.1016/j.cell.2007.11.019

Park, 2008, Reprogramming of human somatic cells to pluripotency with defined factors, Nature, 451, 141, 10.1038/nature06534

Lautenschlaeger, 2012, Endoplasmic reticulum stress and the ER mitochondrial calcium cycle in amyotrophic lateral sclerosis, Amyotroph Lateral Scler, 13, 166, 10.3109/17482968.2011.641569

Hu, 2010, Neural differentiation of human induced pluripotent stem cells follows developmental principles but with variable potency, Proc Natl Acad Sci USA, 107, 4335, 10.1073/pnas.0910012107

Pourquié, 1995, Control of somite patterning by signals from the lateral plate, Proc Natl Acad Sci USA, 92, 3219, 10.1073/pnas.92.8.3219

Parr, 1993, Mouse Wnt genes exhibit discrete domains of expression in the early embryonic CNS and limb buds, Development, 119, 247, 10.1242/dev.119.1.247

Aulehla, 2010, Signaling gradients during paraxial mesoderm development, Cold Spring Harb Perspect Biol, 2, a000869, 10.1101/cshperspect.a000869

Hofmann, 2004, WNT signaling, in synergy with T/TBX6, controls Notch signaling by regulating Dll1 expression in the presomitic mesoderm of mouse embryos, Genes Dev, 18, 2712, 10.1101/gad.1248604

Zhang, 2006, Receptor specificity of the fibroblast growth factor family. The complete mammalian FGF family, J Biol Chem, 281, 15694, 10.1074/jbc.M601252200

Kästner, 2000, Gene expression patterns of the fibroblast growth factors and their receptors during myogenesis of rat satellite cells, J Histochem Cytochem, 48, 1079, 10.1177/002215540004800805

Marics, 2002, FGFR4 signaling is a necessary step in limb muscle differentiation, Development, 129, 4559, 10.1242/dev.129.19.4559

Yu, 2004, Distinct transcriptional control and action of fibroblast growth factor receptor 4 in differentiating skeletal muscle cells, Lab Invest, 84, 1571, 10.1038/labinvest.3700187

Fong, 2012, Genetic and epigenetic determinants of neurogenesis and myogenesis, Dev Cell, 22, 721, 10.1016/j.devcel.2012.01.015

Takemoto, 2011, Tbx6-dependent Sox2 regulation determines neural or mesodermal fate in axial stem cells, Nature, 470, 394, 10.1038/nature09729

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Stem cells translational medicine

Tập 3 Số 5

564-574

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