Fetal tissue engineering: in vitro analysis of muscle constructs |
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Authors: | Fuchs Julie R Pomerantseva Irina Ochoa Erin R Vacanti Joseph P Fauza Dario O |
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Affiliation: | a Harvard Center for Minimally Invasive Surgery, Harvard Medical School; Center for the Integration of Medicine and Innovative Technologies, Massachusetts General Hospital; and Children’s Hospital, Boston, MA, USA |
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Abstract: | Background/purpose: This study was aimed at examining the impact of different tissue engineering techniques on fetal muscle construct architecture.Methods: Myoblasts from ovine specimens of fetal skeletal muscle were expanded in culture and their growth rates determined. Cells were seeded at different densities onto 3 scaffold types, namely polyglycolic acid (PGA) treated with poly-l-lactic acid (PLLA), a composite of PGA with poly-4-hydroxybutyrate (P4HB), and a collagen hydrogel. Constructs were maintained in a bioreactor and submitted to histologic, scanning electron microscopy, and DNA analyses at different time-points. Statistical analysis was by the likelihood ratio and paired Student’s t tests (P < .05).Results: Fetal myoblasts proliferated at faster rates than expected from neonatal cells. Cell attachment was enhanced in the PGA/PLLA matrix and collagen hydrogel when compared with the PGA/P4HB composite. Necrosis was observed at the center of all constructs, directly proportional to cell seeding density and time in the bioreactor.Conclusions: Fetal myoblasts can be expanded rapidly in culture and attach well to PGA/PLLA, as well as collagen hydrogel but less optimally to PGA/P4HB. Excessive cell seeding density and bioreactor time may worsen final construct architecture. These findings should be considered during in vivo trials of muscle replacement by engineered fetal constructs. |
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Keywords: | Tissue engineering fetal muscle myoblast fetal surgery congenital anomalies transplantation |
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