Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Av. E. Mounier 52, Brussels, Belgium.
Pôle de Recherche en Pédiatrie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Av. E. Mounier 52, Brussels, Belgium.
Mol Hum Reprod. 2019 Apr 1;25(4):184-193. doi: 10.1093/molehr/gaz008.
Adipose tissue-derived stem cells (ASCs) have multilineage differentiation potential, proangiogenic properties, and the ability to enhance vascularization in xenografted human ovarian tissue. The aim of the present study was to identify the mechanisms behind the proangiogenic effects of ASCs. For this purpose, severe combined immunodeficient (SCID) mice were grafted with frozen-thawed human ovarian tissue. ASCs were labeled by lentiviral transfection for expression of enhanced green fluorescent protein (eGFP), and human ovarian tissue was grafted using a previously described two-step procedure. In the control group, ovarian tissue was transplanted using the standard one-step approach. Samples were collected and analyzed after 7 days. Detection of the eGFP antigen by immunofluorescence showed ASCs surrounding and infiltrating ovarian tissue grafts. Significantly higher vessel density was observed in the ASC group (P = 0.0182 versus control) on Day 7. Co-expression of eGFP, CD34 and CD31 was demonstrated in human vessels, confirming ASC differentiation into human endothelial cell lineages. Increased gene expression of vascular endothelial growth factor (VEGF) was also shown in the ASC group (P = 0.0182 versus control). Immunohistochemistry targeting anti-human VEGF revealed significantly higher expression levels in the ASC group (P = 0.033 versus control), while VEGF and eGFP immunofluorescence showed greater growth factor expression in areas surrounding ASCs. In conclusion, ASCs differentiate into human vessels and promote secretion of VEGF when transplanted together with human ovarian tissue to SCID mouse peritoneum using a two-step ovarian tissue grafting procedure. This is a promising step towards potentially improving ovarian tissue quality and lifespan. Long-term studies should be conducted to investigate ASC safety and efficacy in the context of ovarian tissue transplantation.
脂肪组织来源的干细胞（ASCs）具有多能分化潜能、促血管生成特性，并能增强异种移植人卵巢组织中的血管生成。本研究旨在探讨 ASCs 促血管生成作用的机制。为此，将冷冻-解冻的人卵巢组织移植到严重联合免疫缺陷（SCID）小鼠体内。通过慢病毒转染对 ASCs 进行标记，以表达增强型绿色荧光蛋白（eGFP），并采用先前描述的两步法移植人卵巢组织。在对照组中，采用标准的一步法移植卵巢组织。在第 7 天收集和分析样本。免疫荧光检测 eGFP 抗原显示 ASCs 环绕和浸润卵巢组织移植物。第 7 天，ASCs 组的血管密度显著高于对照组（P = 0.0182）。在人血管中证明了 eGFP、CD34 和 CD31 的共表达，证实了 ASC 向人内皮细胞谱系的分化。ASCs 组的血管内皮生长因子（VEGF）基因表达也增加（P = 0.0182 与对照组相比）。针对抗人 VEGF 的免疫组织化学显示，ASCs 组的表达水平显著高于对照组（P = 0.033 与对照组相比），而 VEGF 和 eGFP 免疫荧光显示在 ASCs 周围区域有更高的生长因子表达。总之，当通过两步法卵巢组织移植程序将 ASCs 与人类卵巢组织一起移植到 SCID 小鼠的腹膜腔中时，ASCs 会分化为人血管，并促进 VEGF 的分泌。这是潜在改善卵巢组织质量和寿命的有希望的一步。应进行长期研究，以调查 ASC 在卵巢组织移植背景下的安全性和疗效。
