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

基于支架的脂肪组织干细胞在大鼠冻融卵巢自体移植中的输送:在大鼠模型中的初步研究

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作者:Luciana Lamarão Damous, Juliana Sanajotti Nakamuta, Ana Elisa Teofilo Saturi de Carvalho, Kátia Cândido Carvalho, José Maria Soares Jr, Manuel de Jesus Simões, José Eduardo Krieger, Edmund C Baracat

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.

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.

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.

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).

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