Bioprinted Notch ligand to function as stem cell niche improves muscle regeneration in dystrophic muscle

Zewei Sun, Xianlin Yue, Lei Liu, Ying Li, Jie Cui, Dong Li, Lee Weiss, Phil Campbell, Yanling Mu, Johnny Huard, Xiaodong Mu

Article ID: 711
Vol 9, Issue 3, 2023, Article identifier:

VIEWS - 71 (Abstract) 47 (PDF)


In Duchenne muscular dystrophy, dystrophic muscle phenotypes are closely associated with the exhaustion of muscle stem cells. Transplantation of muscle stem cells has been widely studied for improving muscle regeneration, but poor cell survival and self-renewal, rapid loss of stemness, and limited dispersion of grafted cells following transplantation have collectively hindered the overall success of this strategy. Optimized mechanisms for maintaining and improving stem cell function are naturally present in the microenvironment of the stem cell niche in healthy muscles. Therefore, one logical strategy toward improving stem cell function and efficiency of stem cell transplantation in diseased muscles would be the establishment of a microenvironment mimicking some key aspects of healthy native stem cell niches. Here, we applied inkjet-based bioprinting technology to engineer a mimicked artificial stem cell niche in dystrophic muscle, comprising stem cell niche regulating factors (Notch activator DLL1) bioprinted onto 3D DermaMatrix construct. The recombinant DLL1 protein, DLL1 (mouse): Fc (human) (rec), was applied here as the Notch activator. Bioprinted DermaMatrix construct was seeded with muscle stem cells in vitro, and increased stem cell maintenance and repressed myogenic differentiation process was observed. DLL1 bioprinted DermaMatrix construct was then engrafted into dystrophic muscle of mdx/scid mice, and the improved cell engraftment and progression of muscle regeneration was observed 10 days after engraftment. Our results demonstrated that bioprinting of Notch activator within 3D construct can be applied to serve as muscle stem cell niche and improve the efficacy of muscle stem cell transplantation in diseased muscle


Muscle dystrophy, Stem cell niche, Muscle stem cell, Notch signaling

Full Text:

Download PDF

Included Database


Sacco A, Mourkioti F, Tran R, et al., 2010, Short telomeres and stem cell exhaustion model Duchenne muscular dystrophy in mdx/mTR mice. Cell, 143(7):1059–1071.

Jang YC, Sinha M, Cerletti M, et al., 2011, Skeletal muscle stem cells: Effects of aging and metabolism on muscle regenerative function. Cold Spring Harb Symp Quant Biol, 76:101–111.

Usas A, Maciulaitis J, Maciulaitis R, et al., 2011, Skeletal muscle-derived stem cells: Implications for cell-mediated therapies. Medicina (Kaunas), 47(9):469–479.

Ikezawa M, Cao B, Qu Z, et al., 2003, Dystrophin delivery in dystrophin-deficient DMDmdx skeletal muscle by isogenic muscle-derived stem cell transplantation. Hum Gene Ther, 14(16):1535–1546.

Winkler T, von Roth P, Matziolis G, et al., 2009, Dose-response relationship of mesenchymal stem cell transplantation and functional regeneration after severe skeletal muscle injury in rats. Tissue Eng Part A, 15(3):487–492.

Tremblay JP, Malouin F, Roy R, et al., 1993, Results of a triple blind clinical study of myoblast transplantations without immunosuppressive treatment in young boys with Duchenne muscular dystrophy. Cell Transplant, 2(2): 99–112.

Drowley L, Okada M, Beckman S, et al., 2010, Cellular antioxidant levels influence muscle stem cell therapy. Mol Ther, 18(10):1865–1873.

Maclean S, Khan WS, Malik AA, et al., 2012, The potential of stem cells in the treatment of skeletal muscle injury and disease. Stem Cells Int, 2012:282348.

Gussoni E, Blau HM, Kunkel LM, 1997, The fate of individual myoblasts after transplantation into muscles of DMD patients. Nat Med, 3(9):970–977.

Karpati G, Holland P, Worton RG, 1992, Myoblast transfer in DMD: Problems in the interpretation of efficiency. Muscle Nerve, 15(10):1209–1210.

Mayeuf A, Relaix F, 2011, Notch pathway: From development to regeneration of skeletal muscle. Med Sci (Paris), 27(5):521–526.

Carey KA, Farnfield MM, Tarquinio SD, et al., 2007, Impaired expression of Notch signaling genes in aged human skeletal muscle. J Gerontol A Biol Sci Med Sci, 62(1):9–17.

Conboy IM, Rando TA, 2002, The regulation of Notch signaling controls satellite cell activation and cell fate determination in postnatal myogenesis. Dev Cell, 3(3):397–409.

Acharyya S, Sharma SM, Cheng AS, et al., 2010, TNF inhibits Notch-1 in skeletal muscle cells by Ezh2 and DNA methylation mediated repression: Implications in Duchenne muscular dystrophy. PLoS One, 5(8):e12479.

Sweeney C, Morrow D, Birney YA, et al., 2004, Notch 1 and 3 receptor signaling modulates vascular smooth muscle cell growth, apoptosis, and migration via a CBF-1/RBP-Jk dependent pathway. FASEB J, 18(12):1421–1423.

Li Y, Hiroi Y, Liao JK, 2010, Notch signaling as an important mediator of cardiac repair and regeneration after myocardial infarction. Trends Cardiovasc Med, 20(7):228–231.

Carlson ME, Hsu M, Conboy IM, 2008, Imbalance between pSmad3 and Notch induces CDK inhibitors in old muscle stem cells. Nature, 454(7203):528–532.

Yang K, Proweller A, 2011, Vascular smooth muscle Notch signals regulate endothelial cell sensitivity to angiogenic stimulation. J Biol Chem, 286(15):13741–13753.

Phng LK, Gerhardt H, 2009, Angiogenesis: A team effort coordinated by notch. Dev Cell, 16(2):196–208.

Boonen KJ, Post MJ, 2008, The muscle stem cell niche: Regulation of satellite cells during regeneration. Tissue Eng Part B Rev, 14(4):419–431.

Kuang S, Kuroda K, Le Grand F, et al., 2007, Asymmetric self-renewal and commitment of satellite stem cells in muscle. Cell, 129(5):999–1010.

Chakkalakal JV, Jones KM, Basson MA, et al., 2012, The aged niche disrupts muscle stem cell quiescence. Nature, 490(7420):355–360.

Bjornson CR, Cheung TH, Liu L, et al., 2012, Notch signaling is necessary to maintain quiescence in adult muscle stem cells. Stem Cells, 30(2):232–242.

Mourikis P, Sambasivan R, Castel D, et al., 2012, A critical requirement for notch signaling in maintenance of the quiescent skeletal muscle stem cell state. Stem Cells, 30(2):243–252.

Fukada S, Yamaguchi M, Kokubo H, et al., 2011, Hesr1 and Hesr3 are essential to generate undifferentiated quiescent satellite cells and to maintain satellite cell numbers. Development, 138(21):4609–4619.

Brohl D, Vasyutina E, Czajkowski MT, et al., 2012, Colonization of the satellite cell niche by skeletal muscle progenitor cells depends on Notch signals. Dev Cell, 23(3):469–481.

Lepper C, Low S, Partridge TA, 2012, The satellite cell builds its nest under Notch’s guidance. Cell Stem Cell, 11(4):443–444.

Vieira NM, Elvers I, Alexander MS, et al., 2015, Jagged 1 rescues the Duchenne muscular dystrophy phenotype. Cell, 163(5):1204–1213.

Gharaibeh B, Lu A, Tebbets J, et al., 2008, Isolation of a slowly adhering cell fraction containing stem cells from murine skeletal muscle by the preplate technique. Nat Protoc, 3(9):1501–1509.

Ota S, Uehara K, Nozaki M, et al., 2011, Intramuscular transplantation of muscle-derived stem cells accelerates skeletal muscle healing after contusion injury via enhancement of angiogenesis. Am J Sports Med, 39(9):1912– 1922.

Sekiya N, Tobita K, Beckman S, et al., 2013, Muscle-derived stem cell sheets support pump function and prevent cardiac arrhythmias in a model of chronic myocardial infarction. Mol Ther, 21(3):662–9.

Matai I, Kaur G, Seyedsalehi A, et al., 2020, Progress in 3D bioprinting technology for tissue/organ regenerative engineering. Biomaterials, 226:119536.

Vijayavenkataraman S, Yan WC, Lu WF, et al., 2018, 3D bioprinting of tissues and organs for regenerative medicine. Adv Drug Deliv Rev, 132:296–332.

Ng WL, Chua CK, Shen Y-F, 2019, Print me an organ! Why we are not there yet. Progr Polym Sci, 97:101145.

Ng WL, Lee JM, Zhou M, et al., 2020, Vat polymerization-based bioprinting-process, materials, applications and regulatory challenges. Biofabrication, 12(2):022001.

Ng WL, Huang X, Shkolnikov V, et al., 2022, Controlling droplet impact velocity and droplet volume: Key factors to achieving high cell viability in sub-nanoliter droplet-based bioprinting. Int J Bioprint, 8(1):424.

Zhou C, Yang Y, Wang J, et al., 2021, Ferromagnetic soft catheter robots for minimally invasive bioprinting. Nat Commun, 12(1):5072.

Jiang T, Munguia-Lopez GJ, Flores-Torres S, et al., 2019, Extrusion bioprinting of soft materials: An emerging technique for biological model fabrication. Appl Phys Rev, 6(1):011310., 10.1063/1.5085013, 10.1063/1.5055659, 10.1063/1.5053909

Eisenberg MC, Kim Y, Li R, et al., 2011, Mechanistic modeling of the effects of myoferlin on tumor cell invasion. Proc Natl Acad Sci U S A, 108(50):20078–20083.

Yerneni SS, Whiteside TL, Weiss LE, et al., 2019, Bioprinting exosome-like extracellular vesicle microenvironments. Bioprinting, 13:e00041.

Cooper GM, Miller ED, DeCesare GE, et al., 2010, Inkjet-based biopatterning of bone morphogenetic protein-2 to spatially control calvarial bone formation. Tissue Eng Part A, 16:1749–1759.

Ker ED, Nain AS, Weiss LE, et al., 2011, Bioprinting of growth factors onto aligned sub-micron fibrous scaffolds for simultaneous control of cell differentiation and alignment. Biomaterials, 32(32):8097–8107.

Herberg S, Kondrikova G, Periyasamy-Thandavan S, et al., 2014, Inkjet-based biopatterning of SDF-1beta augments BMP-2-induced repair of critical size calvarial bone defects in mice. Bone, 67:95–103.

Skuk D, Tremblay JP, 2000, Progress in myoblast transplantation: A potential treatment of dystrophies. Microsc Res Tech, 48(3-4):213–222. 3/4<213::AID-JEMT9>3.0.CO;2-Z

Bouchentouf M, Benabdallah BF, Tremblay JP, 2004, Myoblast survival enhancement and transplantation success improvement by heat-shock treatment in mdx mice. Transplantation, 77(9):1349–1356.

Skuk D, Caron NJ, Goulet M, et al., 2003, Resetting the problem of cell death following muscle-derived cell transplantation: Detection, dynamics and mechanisms. J Neuropathol Exp Neurol, 62(9):951–967.

Qu-Petersen Z, Deasy B, Jankowski R, et al., 2002, Identification of a novel population of muscle stem cells in mice: Potential for muscle regeneration. J Cell Biol, 157(5):851–864.

Deasy BM, Jankowski RJ, Huard J, 2001, Muscle-derived stem cells: Characterization and potential for cell-mediated therapy. Blood Cells Mol Dis, 27(5):924–933.

Parker MH, Loretz C, Tyler AE, et al., 2012, Activation of Notch signaling during ex vivo expansion maintains donor muscle cell engraftment. Stem Cells, 30(10):2212–2220.



  • There are currently no refbacks.

Copyright (c) 2023 Author(s).

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.