title: Scaffold-based delivery of adipose tissue-derived stem cells in rat frozen-thawed ovarian autografts: preliminary studies in a rat model.
abstract: This study aimed to evaluate whether a gelatin-based Gelfoam sponge is feasible as a scaffold for adipose tissue-derived stem cell (ASC) therapy in rat frozen-thawed ovarian autografts.
  
  
  
  Two sets of studies were performed. The in vitro set evaluated ASCs' viability in the Gelfoam scaffold at different times of co-culturing (after 24, 48, 72, 96, and 120 h). The in vivo set used 20 12-week-old adult female Wistar rats. Frozen-thawed ovarian grafts were treated with ASCs delivered in Gelfoam scaffolds immediately after an autologous retroperitoneal transplant (ASCs-GS, n = 10). The controls received Gelfoam with a culture medium (GS, n = 10). Assessment of graft quality was conducted by vaginal smears (until euthanasia on the 30th postoperative day), histological analyses, follicular density, and viability and fibrosis. Immunohistochemical staining for VEGF-A expression, vascular network (vWF), apoptosis (caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)), cell proliferation (Ki-67), and hormone receptors (estrogen and progesterone) were performed.
  
  
  
  The cells remained viable in Gelfoam for up to 120 h of co-culturing. The graft morphology was similar among the groups. ASC therapy promoted the earlier resumption of the estrous phase (GS 16.6 ± 3 vs. ASCs-GS 12.8 ± 1.3 days) and enhanced estrogen receptors compared with the controls (p < 0.05) without interfering with the quantity and viability of the ovarian follicles, fibrosis, endothelial cells, VEGF immunoexpression, apoptosis, or cell proliferation (p > 0.05).
  
  
  
  The Gelfoam scaffold could be a feasible and safe non-invasive technique for ASC delivery in the treatment of frozen-thawed ovarian autografts. Future studies should evaluate the real benefit of this treatment on the survival and endocrine activity of the graft.
authors.affiliation: Gynecology Discipline, Laboratory of Structural and Molecular Gynecology (LIM-58), Faculdade de Medicina da Universidade de São Paulo, Dr Arnaldo av 455, 2nd floor, room 2113, Pacaembu, São Paulo, Brazil, 01246-903, lucianadamous@gmail.com.
authors.forename: Luciana Lamarão
authors.initials: LL
authors.lastname: Damous
authors.forename: Juliana Sanajotti
authors.initials: JS
authors.lastname: Nakamuta
authors.forename: Ana Elisa Teofilo
authors.initials: AE
authors.lastname: Saturi de Carvalho
authors.forename: Kátia Cândido
authors.initials: KC
authors.lastname: Carvalho
authors.forename: José Maria
authors.initials: JM
authors.lastname: Soares
authors.forename: Manuel de Jesus
authors.initials: Mde J
authors.lastname: Simões
authors.forename: José Eduardo
authors.initials: JE
authors.lastname: Krieger
authors.forename: Edmund C
authors.initials: EC
authors.lastname: Baracat
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D017209:Apoptosis; D064592:Autografts; D049109:Cell Proliferation; D002470:Cell Survival; D018920:Coculture Techniques; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D005780:Gelatin; D018919:Neovascularization, Physiologic; D006080:Ovarian Follicle / Q000502:physiology; D010053:Ovary / Q000166:cytology / Q000502:physiology / Q000637:transplantation*; D017208:Rats, Wistar; D033581:Stem Cell Transplantation / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D005780:Gelatin