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

  • Fertility Preservation
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Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

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.

Methods

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.

Results

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).

Conclusion

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.

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Acknowledgments

We thank São Paulo Research Foundation (FAPESP, Pio XI St, 1500, Alto da Lapa, São Paulo/SP-Brazil, 05468-901) for grant support (process numbers: 2010/17897-5 and 2012/09469-9) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for post-doctoral scholarship; Esmeralda Miristene Eher, Angela Batista Santos, Maria Cristina Rodrigues Medeiros, and Sandra de Moraes Fernezlian (Immunohistochemistry Laboratory/Pathology Department/FMUSP) for technical assistance with immunohistochemistry assay; Leonard Medeiros da Silva (pathologist) for the help in the analysis of morphological data; and American Expert Journal for the english editing service.

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Authors and Affiliations

  1. 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

    Luciana Lamarão Damous, Kátia Cândido Carvalho, José Maria Soares-Jr & Edmund C. Baracat

  2. Laboratory of Genetics and Molecular Cardiology, Heart Institute (Incor), Faculdade de Medicina da Universidade de São Paulo, Dr Enéas de Carvalho Aguiar Av 44, 10th floor, Cerqueira Cesar, São Paulo, Brazil, 05403-000

    Juliana Sanajotti Nakamuta, Ana Elisa Teofilo Saturi de Carvalho & José Eduardo Krieger

  3. Department of Morphology and Genetics, Universidade Federal de São Paulo (UNIFESP), Botucatu St 740. Ed. Lemos Torres, 2nd floor, Vila Clementino, São Paulo, Brazil, 04023-009

    Manuel de Jesus Simões

  4. Galvão Bueno St, 499. Bloco A. Apto31, Liberdade, São Paulo, Brazil, 01506-000

    Luciana Lamarão Damous

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  1. Luciana Lamarão Damous
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Correspondence to Luciana Lamarão Damous.

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Capsule The Gelfoam scaffold could be a feasible and safe non-invasive technique for ASC delivery in the treatment of frozen-thawed ovarian autografts.

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Damous, L.L., Nakamuta, J.S., Saturi de Carvalho, A.E.T. et al. Scaffold-based delivery of adipose tissue-derived stem cells in rat frozen-thawed ovarian autografts: preliminary studies in a rat model. J Assist Reprod Genet 32, 1285–1294 (2015). https://doi.org/10.1007/s10815-015-0527-x

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  • DOI: https://doi.org/10.1007/s10815-015-0527-x

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