Details

Autor(en) / Beteiligte

• Janet Shansky
• Michael Del Tatto
• Joseph Chromiak
• Vandenburgh, Herman

Titel

A Simplified Method for Tissue Engineering Skeletal Muscle Organoids in vitro

Ist Teil von

• In vitro cellular & developmental biology. Animal, 1997-10, Vol.33 (9), p.659-661

Ort / Verlag

Germany: Society for In Vitro Biology

Erscheinungsjahr

1997

Quelle

SpringerLINK Contemporary (Konsortium Baden-Württemberg)

Beschreibungen/Notizen

• Tissue-engineered three-dimensional skeletal muscle organ-like structures have been formed in vitro from primary myoblasts by several different techniques. The resulting "organoids" display many of the characteristics of in vivo muscle including parallel arrays of postmitotic fibers organized into fascicle-like structures with tendon-like ends. They are contractile, express adult isoforms of contractile proteins, can perform directed work, and can be maintained in culture for at least 4 wk. The in vivo-like characteristics and durability of the muscle organoids make them an ideal model system for long-term in vitro studies on mechanotransduction mechanisms and on muscle atrophy induced by decreased muscle tension. Tissue-engineered skeletal muscle organoids have also been used as an implantable device for the systemic delivery of recombinant proteins. Current methods for muscle organoid formation are limited by the number which can be formed at one time and by the complicated procedures necessary to induce organogenesis. Induction of muscle organogenesis from skeletal myoblasts utilizes either internally or externally generated mechanical tension as an important element for the alignment of the fusing myoblasts and subsequent organoid formation. For example, a computerized mechanical cell stimulator device provides external longitudinal mechanical forces during the first 2-3 d of myoblast proliferation to orient the developing myofibers parallel to each other and to the direction of stretch. After 6-10 d of additional complex mechanical stimulation, the edges of the cell monolayer roll in to form an organ-like structure. Similar muscle organogenesis has been induced passively when high density monolayers of avian myoblasts are plated onto a collagen-coated, flexible plastic substrate held in place by stainless steel pins. After 10-15 d of culture, the differentiated myofibers begin twitching spontaneously, generating internal tensions which cause the cell sheet to detach from the flexible substrate while remaining held in place by the stainless steel pins. Although these methods have been useful for small-scale muscle organogenesis studies, they have not been scaled up to provide the large number of samples necessary for more complex studies. In this report, we describe a simple method for generating large numbers of muscle organoids from either primary embryonic avian or neonatal rodent myoblasts.