Three-dimensional culture in a bioengineered matrix and somatic cell complementation to improve growth and survival of bovine preantral follicles.

IF 3.2 3区医学 Q2 GENETICS & HEREDITY

Journal of Assisted Reproduction and Genetics Pub Date : 2025-05-01 Epub Date: 2025-05-20 DOI:10.1007/s10815-025-03497-3

Juliana I Candelaria, Ramon C Botigelli, Carly Guiltinan, Ariella Shikanov, Anna C Denicol

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引用次数: 0

Abstract

Purpose: Here, we explored poly(ethylene glycol) (PEG) bioengineered hydrogels for bovine preantral follicle culture with or without ovarian cell co-culture and examined the potential for differentiation of bovine embryonic stem cells (bESCs) towards gonadal somatic cells to develop a system better mimicking the ovarian microenvironment.

Methods: Bovine preantral follicles were first cultured in two-dimensional (2D) control or within PEG hydrogels (3D) and then co-cultured within PEG hydrogels with bovine ovarian cells (BOCs) to determine growth and viability. Finally, we tested conditions to drive differentiation of bESCs towards the intermediate mesoderm and bipotential gonad fate.

Results: Primary follicles grew over the 10-day culture period in PEG hydrogels compared to 2D control. Early secondary follicles maintained a similar diameter within the PEG while control follicles decreased in size. Follicles lost viability after co-encapsulation with BOCs; BOCs lost stromal cell signature over the culture period within hydrogels. Induction of bESCs towards gonadal somatic fate under WNT signaling was sufficient to upregulate intermediate mesoderm (LHX1) and early coelomic epithelium/bipotential gonad markers (OSR1, GATA4, WT1). Higher BMP4 concentrations upregulated the lateral plate mesoderm marker FOXF1. PAX3 expression was not induced, indicating absence of the paraxial mesoderm lineage.

Conclusions: Culture of primary stage preantral follicles in PEG hydrogels promoted growth compared to controls; BOCs did not maintain identity in the PEG hydrogels. Collectively, we demonstrate that PEG hydrogels can be a potential culture system for early preantral follicles pending refinements, which could include addition of ESC-derived ovarian somatic cells using the protocol described here.

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生物工程基质三维培养和体细胞互补提高牛腔前卵泡的生长和存活。

目的：在此，我们探索了聚乙二醇（PEG）生物工程水凝胶用于与卵巢细胞共培养或不共培养的牛腔前卵泡培养，并研究了牛胚胎干细胞（bESCs）向性腺体细胞分化的潜力，以建立一个更好地模拟卵巢微环境的系统。方法：先在二维对照（2D）或聚乙二醇水凝胶（3D）中培养牛腔前卵泡，然后在聚乙二醇水凝胶中与牛卵巢细胞（BOCs）共培养，测定其生长和活力。最后，我们测试了驱动bscs向中胚层和双潜能性腺分化的条件。结果：与2D对照相比，PEG水凝胶的初代卵泡生长时间超过10天。早期次生卵泡在PEG内保持相似的直径，而对照卵泡的大小减小。与BOCs共包埋后卵泡丧失活力；BOCs在水凝胶内的培养过程中失去了基质细胞特征。在WNT信号下，bscs对性腺体细胞命运的诱导足以上调中胚层（LHX1）和早期体腔上皮/双电位性腺标志物（OSR1, GATA4, WT1）。较高的BMP4浓度上调了侧板中胚层标志物FOXF1。PAX3未被诱导表达，表明不存在近轴中胚层谱系。结论：与对照组相比，PEG水凝胶培养初生期腔前卵泡促进了卵泡的生长；BOCs在PEG水凝胶中不保持身份。总的来说，我们证明了PEG水凝胶可以成为一种潜在的早期腔前卵泡培养系统，有待改进，其中可能包括使用本文描述的方案添加esc衍生的卵巢体细胞。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Assisted Reproduction and Genetics 医学-妇产科学

CiteScore

5.70

自引率

9.70%

发文量

286

审稿时长

1 months

期刊介绍： The Journal of Assisted Reproduction and Genetics publishes cellular, molecular, genetic, and epigenetic discoveries advancing our understanding of the biology and underlying mechanisms from gametogenesis to offspring health. Special emphasis is placed on the practice and evolution of assisted reproduction technologies (ARTs) with reference to the diagnosis and management of diseases affecting fertility. Our goal is to educate our readership in the translation of basic and clinical discoveries made from human or relevant animal models to the safe and efficacious practice of human ARTs. The scientific rigor and ethical standards embraced by the JARG editorial team ensures a broad international base of expertise guiding the marriage of contemporary clinical research paradigms with basic science discovery. JARG publishes original papers, minireviews, case reports, and opinion pieces often combined into special topic issues that will educate clinicians and scientists with interests in the mechanisms of human development that bear on the treatment of infertility and emerging innovations in human ARTs. The guiding principles of male and female reproductive health impacting pre- and post-conceptional viability and developmental potential are emphasized within the purview of human reproductive health in current and future generations of our species. The journal is published in cooperation with the American Society for Reproductive Medicine, an organization of more than 8,000 physicians, researchers, nurses, technicians and other professionals dedicated to advancing knowledge and expertise in reproductive biology.