Gene expression profile in experimental frozen-thawed ovarian grafts treated with scaffold-base delivery of adipose tissue-derived stem cells

• Published in: Clinics (São Paulo, Brazil) Vol. 77; p. 100066
• Main Authors: Damous, Luciana Lamarão, Shiroma, Marcos Eiji, Carvalho, Ana Elisa Teófilo Saturi de, Soares-Jr, José Maria, Krieger, José Eduardo, Baracat, Edmund C.
• Format: Journal Article
• Language: English
• Published: Elsevier España, S.L.U 01.01.2022; Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo; Faculdade de Medicina / USP
• Subjects: Fertility preservation; Gene expression; MEDICINE, GENERAL & INTERNAL; Original; Ovarian tissue transplantation; Rat; Stem cells; clade B (ovalbumin); Ovarian tissue transplantation; Rat; RNA; DMSO; Qpcr; G1; G2; rASCs; DMEM; SD; Serpinb5; RT-PCR; LTa; GC; MSC; Cxcl; Casp14; PBS; Fertility preservation; IL; large; VEGF; POI; Gene expression; Hprt1; Actb, Actin; MMP; Ldha; TNF-β; Stem cells; Rplp1; ANG; HE; B2m
• ISSN: 1807-5932; 1980-5322; 1980-5322
• DOI: 10.1016/j.clinsp.2022.100066

•The scaffold-based delivery therapy with adipose tissue-derived stem cells in the rat ovarian autografts seems to be the best option when compared to direct injection or systemic route.•Ovarian grafts treated with adipose tissue-derived stem cells showed the highest number of genes over-regulated in the apoptosis pathway, compared to inflammation cytokines and angiogenesis pathway.•Capsase14 was the most over-regulated gene in the apoptosis pathway.•The treatment with adipose tissue-derived stem cells in ovarian grafts treated didn't compromise the ovary graft morphology and viability for short time. Gelfoam scaffold is a feasible and safe non-invasive technique for Adipose tissue-derived Stem Cell (ASC)-delivery in the treatment of frozen-thawed ovarian autografts. This study seeks to analyze the genes expression profile of rat frozen-thawed ovarian autografts treated with scaffold-based delivery of adipose tissue-derived stem cells. Eighteen adult Wistar rats were distributed into three groups: Control (frozen-thawed only); Group 1 (G1) and Group 2 (G2) (frozen-thawed ovaries treated with culture medium or ASC, respectively). Both treatments were performed immediately after autologous retroperitoneal transplant with scaffold-based delivery. The ovarian grafts were retrieved 30 days after transplantation. Quantitative gene expression (qPCR) for apoptosis, angiogenesis, and inflammatory cytokines (84 genes in each pathway) were evaluated by RT-PCR. Graft morphology (HE), apoptosis (cleaved-caspase-3), neoangiogenesis (VEGF), and cellular proliferation (Ki-67) were assessed. In grafts treated with ASC, the apoptosis pathway showed the highest number of genes over-regulated – 49 genes – compared to inflammation cytokines and angiogenesis pathway – 36 and 23 genes respectively, compared to grafts treated with culture medium. Serpinb5 family was highlighted in the angiogenesis pathway and Cxcl6 in the inflammation cytokines pathway. In the apoptosis pathway, the most over-regulated gene was Capsase14. ASC treatment promoted the reduction of cleaved caspase-3 in the theca internal layer and increased cell proliferation by Ki-67 in the granulosa layer without altering VEGF. A mild inflammatory infiltrate was observed in both groups. ASC therapy in rat frozen-thawed ovarian autografts promoted an abundance of genes involved with apoptosis and inflammatory cytokines without compromising the ovary graft morphology and viability for short time. Further studies are necessary to evaluate the repercussion of apoptosis and inflammation on the graft in the long term.