Scaffold-based delivery of adipose tissue-derived stem cells in rat frozen-thawed ovarian autografts: preliminary studies in a rat model

Author(s)

Damous, Luciana Lamarao ^[1] ; Nakamuta, Juliana Sanajotti ^[2] ; Teofilo Saturi de Carvalho, Ana Elisa ^[2] ; Carvalho, Katia Candido ^[1] ; Soares-, Jr., Jose Maria ^[1] ; Simoes, Manuel de Jesus ^[3] ; Krieger, Jose Eduardo ^[2] ; Baracat, Edmund C. ^[1]

Affiliation

^[1] Univ Sao Paulo, Fac Med, Lab Struct & Mol Gynecol LIM 58, Gynecol Discipline, BR-01246903 Sao Paulo - Brazil

^[2] Univ Sao Paulo, Fac Med, Heart Inst Incor, Lab Genet & Mol Cardiol, BR-05403000 Sao Paulo - Brazil

^[3] Univ Fed Sao Paulo, Dept Morphol & Genet, BR-04023009 Sao Paulo - Brazil

• Document type: Journal article
• Source: JOURNAL OF ASSISTED REPRODUCTION AND GENETICS; v. 32, n. 8, p. 1285-1294, AUG 2015.
• Web of Science Citations: 3

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. (AU)

• FAPESP's process: 12/09469-9 - Cellular and gene therapy in experimental fresh and cryopreserved ovary transplant
• Grantee: Edmund Chada Baracat
• Support Opportunities: Regular Research Grants
• FAPESP's process: 10/17897-5 - Cellular and gene therapy in experimental fresh and cryopreserved ovary transplant
• Grantee: Luciana Lamarão Damous
• Support Opportunities: Scholarships in Brazil - Post-Doctoral