Capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro.
Capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro.
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DOI：
10.1038/s41598-021-92036-y
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发表时间：
2021-06-15
影响因子：
4.6
通讯作者：
Shikanov A
中科院分区：
文献类型：
--
作者：
Jones A;Bernabé BP;Padmanabhan V;Li J;Shikanov A
体外卵泡培养是一个活跃的研究领域，为儿童癌症幸存者提供生育选择。晚期小鼠卵泡（多层次级和向前）可以成功培养至成熟，以获得减数分裂能力的卵母细胞用于受精，但原始和初级卵泡通常在培养中死亡，因为早期卵泡发育的许多关键成分仍然未知，难以在体外模拟。为了设计一个高效、可重复的仿生三维培养系统，必须揭示早期卵泡发生的详细机制。以往的研究表明，与单独培养的单个卵泡相比，小鼠原代卵泡群共培养可以达到排卵前的大小并产生有能力的卵母细胞，但卵泡共培养协同作用的因素仍然未知。为了探索成功的卵泡共培养的潜在机制，我们对封装在0.3%藻酸盐水凝胶中的小鼠卵泡进行了时间过程实验，并在两种条件下进行了比较：5个组（5X）与10个组（10X）。在12天的过程中，每2天分离卵泡并分离体细胞用于基于微阵列的基因表达分析（对于5X，n = 380个卵泡，对于10X，n = 430个卵泡）。与较小队列（5X）中共培养的卵泡细胞相比，较大组（10X）中共培养的卵泡中的基因活性具有关键基因和途径（如催乳素信号传导和血管生成相关基因）的不同转录组学特征。为了对培养中生长的卵泡的结果进行基准测试，我们将我们的微阵列数据与Bernabé等人先前发表的从卵巢中新鲜分离的小鼠卵泡的数据进行了比较。这些数据集的比较确定了天然微环境和工程体外系统中卵泡发生之间的相似性和差异。对体外卵泡生长的更详细了解不仅可以提供更好的培养方法，还可以推动该领域为儿童癌症幸存者提供更好的生育选择。
In vitro ovarian follicle culture is an active area of research towards providing fertility options for survivors of childhood cancer. Late-stage murine follicles (multilayer secondary and onwards) can be cultured successfully to maturity to obtain a meiotically competent oocyte for fertilization, but primordial and primary follicles usually die in culture because many key components of early follicle development are still unknown and difficult to mimic in vitro. To engineer a biomimetic three-dimensional culture system with high efficacy and reproducibility for the clinic, detailed mechanisms of early folliculogenesis must be uncovered. Previous studies have shown that primary murine follicles co-cultured in groups, in contrast to single follicles cultured in isolation, can reach preovulatory size and produce competent oocytes, but the factors accounting for the synergy of follicle co-culture are still unknown. To probe the underlying mechanisms of successful follicle co-culture, we conducted a time-course experiment for murine follicles encapsulated in 0.3% alginate hydrogels and compared between two conditions: groups of 5 (5X) versus groups of 10 (10X). For every 2 days during the course of 12 days, follicles were dissociated and somatic cells were isolated for microarray-based gene expression analysis (n = 380 follicles for 5X and n = 430 follicles for 10X). Gene activities in follicles co-cultured in larger groups (10X) had a distinct transcriptomic profile of key genes and pathways such as prolactin signaling and angiogenesis-related genes when compared to cells from follicles co-cultured in the smaller cohort (5X). To benchmark the results for follicles grown in culture, we compared our microarray data to data from murine follicles freshly isolated from the ovary at comparable stages of development previously published by Bernabé et al. Comparison of these datasets identified similarities and differences between folliculogenesis in the native microenvironment and the engineered in vitro system. A more detailed understanding of follicle growth in vitro will not only allow for better culture methods but also advance the field towards providing improved fertility options for survivors of childhood cancer.
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影响因子：
6.7
作者：
Hoang, Yvonne D.;McTavish, Kirsten J.;Chang, R. Jeffrey;Shimasaki, Shunichi
通讯作者：
Shimasaki, Shunichi
影响因子：
4.3
作者：
Eden E;Lipson D;Yogev S;Yakhini Z
通讯作者：
Yakhini Z
DOI：
10.1111/asj.12200
发表时间：
2014-06
期刊：
Animal science journal = Nihon chikusan Gakkaiho
影响因子：
--
作者：
Emori C;Sugiura K
通讯作者：
Sugiura K
影响因子：
6.1
作者：
Carlsson, I. B.;Scott, J. E.;Hovatta, O.
通讯作者：
Hovatta, O.
影响因子：
3.7
作者：
Hatzirodos N;Irving-Rodgers HF;Hummitzsch K;Rodgers RJ
通讯作者：
Rodgers RJ