Contractile Skeletal Muscle Tissue-Engineered on an Acellular Scaffold

Abstract

For the reconstructive surgeon, tissue-engineered skeletal muscle may offer reduced donor-site morbidity and an unlimited supply of tissue. Using an acellularized mouse extensor digitorum longus muscle as a scaffold, the authors produced engineered skeletal muscle capable of generating longitudinal force. Eight extensor digitorum longus muscles from adult mice were made acellular using a protocol developed in the authors' laboratory. The acellular muscles were then placed in a bath of 20% fetal bovine serum in Dulbecco's modified Eagle's medium and 100 U/ml penicillin for 1 week at room temperature. C2C12 myoblasts were injected into the acellular muscle matrix using a 26-gauge needle and a 100-microl syringe. The resulting constructs were placed in growth medium for 1 week at 37 degrees C under 5% carbon dioxide, with media changes every 48 hours. The constructs were then placed in differentiation medium for 1 week, with media changes every 48 hours. Isometric contractile force testing of the constructs demonstrated production of longitudinal contractile force on electrical stimulation. A length-tension, or Starling, relationship was observed. Light and electron microscopy studies demonstrated recapitulation of some of the normal histologic features of developing skeletal muscle.

Injectable biomimetic liquid crystalline scaffolds enhance muscle stem cell transplantation

“…Compared with decellularized muscle (62), other natural biomaterials (63,64), or natural/synthetic composites (65), our all-synthetic biomimetic scaffolds allow the transplantation of muscle stem and progenitor cells in a chemically defined environment. Moreover, their neutral pH and instantaneous gelation upon injection in vivo make our biomimetic scaffolds ideal for cell transplantation.…”

Current opportunities and challenges in skeletal muscle tissue engineering

“…Therefore, frequently cell lines are used such as C2C12, which are satellite cells of C3H mice (Dennis et al, 2001;Borschel et al, 2004;Levenberg et al, 2005;Riboldi et al, 2005;Huang et al, 2006a;Boontheekul et al, 2007;Matsumoto et al, 2007). Also, satellite cells harvested from the soleus muscle of rats are used for TE of muscle tissue (Dennis and Kosnik, 2000;Beier et al, 2004;Stern-Straeter et al, 2005;Das et al, 2006;Borschel et al, 2006;Larkin et al, 2006;Bach et al, 2006;Huang et al, 2006b;Boontheekul et al, 2007).…”

“…Although several studies have addressed the development of an optimal non-biodegradable scaffold, such as Shah et al, 2005Sinanan et al, 2004Stern-Straeter et al, 2008Alessandri et al, 2004 Human Brachioradialis muscle Satellite cell Lewis et al, 2000 C3H mice C2C12 Cell line Borschel et al, 2004Dennis et al, 2000Huang et al, 2005Levenberg et al, 2005Riboldi et al, 2005Boontheekul et al, 2007Borschel et al, 2004 Rat Soleus muscle Satellite cell Larkin et al, 2006Dennis et al, 2000Huang et al, 2005Das et al, 2006Beier et al, 2004Bach et al, 2003Stern-Staeter et al, 2008Boontheekul et al, 2007Huang et al, 2005 Rat Tibialis anterior muscle Satellite cell Dennis et al, 2000Boontheekul et al, 2007Kamelger et al, 2004 Rat Latissimus dorsi muscle Satellite cell Kamelger et al, 2004 Rat phosphate-based glass (Shah et al, 2005), biodegradable scaffolds are preferred because, upon degradation, remodelling to the natural muscular ECM can occur. Both synthetic and natural scaffolds have been developed.…”

“…A natural biodegradable 3D scaffold can also be made of acellular muscle ECM (Borschel et al, 2004). A drawback of this type of scaffold is its extreme fragility and the associated difficulties of handling.…”

Decellularization of Human Lip

“…It muscle fibres have a unique orientation and are suspended from the surrounding muscles without any bony attachments. Decellularized muscle seeded with myoblasts has been shown to be capable of generating a contractile force on electrical stimulation [17]. However decellularization of muscle has so far been limited to very small muscles or fragments of muscle and in vivo successes are limited to small animal models [18][19][20][21][22].…”