title: Comparisons of human amniotic mesenchymal stem cell viability in FDA-approved collagen-based scaffolds: Implications for engineered diaphragmatic replacement.
abstract: OBJECTIVE
  We sought to examine amniotic fluid mesenchymal stem cell (afMSC) viability within two FDA-approved collagen-based scaffolds, as a prerequisite to clinical translation of afMSC-based engineered diaphragmatic repair.
  
  
  METHODS
  Human afMSCs were seeded in a human-derived collagen hydrogel and in a bovine-derived collagen sheet at 3 matching densities. Cell viability was analyzed at 1, 3, and 5days using an ATP-based 3D bioluminescence assay. Statistical comparisons were by ANOVA (P<0.05).
  
  
  RESULTS
  There was a highly significant 3-way interaction between scaffold type, seeding density, and time in 3D culture as determinants of cell viability, clearly favoring the human hydrogel (P<0.001). In both scaffolds, cell viability was highest at the highest seeding density of 150,000 cells/mL. Time in 3D culture impacted cell viability at the optimal seeding density in the human hydrogel, with the highest levels on days 1 (P<0.001) and 5 (P=0.05) with no significant effect in the bovine sheet (P=0.39-0.96).
  
  
  CONCLUSIONS
  Among clinically-approved cell delivery vehicles, mesenchymal stem cell viability is significantly enhanced in a collagen hydrogel when compared with a collagen sheet. Cell viability can be further optimized by seeding density and time in 3D culture. These data further support the regulatory viability of clinical trials of engineered diaphragmatic repair.
  
  
  METHODS
  N/A (animal and laboratory study).
authors.affiliation: Department of Surgery, Boston Children's Hospital and Harvard Medical School, Boston, MA, United States.
authors.forename: Hester F
authors.initials: HF
authors.lastname: Shieh
authors.affiliation: Department of Surgery, Boston Children's Hospital and Harvard Medical School, Boston, MA, United States.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Graham
authors.affiliation: Department of Surgery, Boston Children's Hospital and Harvard Medical School, Boston, MA, United States.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Brazzo
authors.affiliation: Department of Surgery, Boston Children's Hospital and Harvard Medical School, Boston, MA, United States.
authors.forename: David
authors.initials: D
authors.lastname: Zurakowski
authors.affiliation: Department of Surgery, Boston Children's Hospital and Harvard Medical School, Boston, MA, United States. Electronic address: dario.fauza@childrens.harvard.edu.
authors.forename: Dario O
authors.initials: DO
authors.lastname: Fauza
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques; D002470:Cell Survival; D003094:Collagen; D065630:Hernias, Diaphragmatic, Congenital / Q000601:surgery*; D059685:Herniorrhaphy / Q000379:methods*; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D003094:Collagen
keywords: Amniotic fluid mesenchymal stem cells; Collagen scaffolds; Congenital diaphragmatic hernia; Diaphragmatic tissue engineering