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Tissue-engineered human bioartificial muscles expressing a foreign recombinant protein for gene therapyMurine skeletal muscle cells transduced with foreign genes and tissue engineered in vitro into bioartificial muscles (BAMs) are capable of long-term delivery of soluble growth factors when implanted into syngeneic mice (Vandenburgh et al., 1996b). With the goal of developing a therapeutic cell-based protein delivery system for humans, similar genetic tissue-engineering techniques were designed for human skeletal muscle stem cells. Stem cell myoblasts were isolated, cloned, and expanded in vitro from biopsied healthy adult (mean age, 42 +/- 2 years), and elderly congestive heart failure patient (mean age, 76 +/- 1 years) skeletal muscle. Total cell yield varied widely between biopsies (50 to 672 per 100 mg of tissue, N = 10), but was not significantly different between the two patient groups. Percent myoblasts per biopsy (73 +/- 6%), number of myoblast doublings prior to senescence in vitro (37 +/- 2), and myoblast doubling time (27 +/- 1 hr) were also not significantly different between the two patient groups. Fusion kinetics of the myoblasts were similar for the two groups after 20-22 doublings (74 +/- 2% myoblast fusion) when the biopsy samples had been expanded to 1 to 2 billion muscle cells, a number acceptable for human gene therapy use. The myoblasts from the two groups could be equally transduced ex vivo with replication-deficient retroviral expression vectors to secrete 0.5 to 2 microg of a foreign protein (recombinant human growth hormone, rhGH)/10(6) cells/day, and tissue engineered into human BAMs containing parallel arrays of differentiated, postmitotic myofibers. This work suggests that autologous human skeletal myoblasts from a potential patient population can be isolated, genetically modified to secrete foreign proteins, and tissue engineered into implantable living protein secretory devices for therapeutic use.
Document ID
20040142026
Document Type
Reprint (Version printed in journal)
Authors
Powell, C. (Brown University Providence, RI 02912, United States)
Shansky, J.
Del Tatto, M.
Forman, D. E.
Hennessey, J.
Sullivan, K.
Zielinski, B. A.
Vandenburgh, H. H.
Date Acquired
August 22, 2013
Publication Date
March 1, 1999
Publication Information
Publication: Human gene therapy
Volume: 10
Issue: 4
ISSN: 1043-0342
Subject Category
Life Sciences (General)
Funding Number(s)
CONTRACT_GRANT: AG14958
CONTRACT_GRANT: HL60502
CONTRACT_GRANT: AG15415
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Musculoskeletal