Spatio-temporal remodelling of the composition and architecture of the human ovarian cortical extracellular matrix during in vitro culture.
Spatio-temporal remodelling of the composition and architecture of the human ovarian cortical extracellular matrix during in vitro culture.
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DOI：
10.1093/humrep/dead008
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发表时间：
2023-03-01
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体外培养如何改变人卵巢皮质细胞外基质（ECM）网络结构？卵巢皮层不同层的ECM组成和结构不同，并在体外培养过程中被重塑。卵巢外基质是组织卵泡和基质细胞的支架。它的组成和结构特性不断演变，以适应卵泡的发育和扩张。原生ECM内原始卵泡培养的组织准备涉及机械松动；这在ECM网络中诱导了未定义的修饰，并改变了细胞间的接触，导致自发的卵泡激活。对6例28-38岁（平均±SD: 32.7±4.1岁）择期剖宫产的女性进行新鲜卵巢皮质活检。将活组织切片切成约4 × 1 × 1 mm的碎片，培养0、2、4或6天(D)。利用组织学和免疫组织学分析，在每个时间点对整个皮质的原始滤泡激活、基质细胞密度和ecm相关蛋白（胶原、弹性蛋白、纤维连接蛋白、层粘连蛋白）阳性面积进行量化。在人卵巢皮层的每一层，即外皮层、中皮层和皮层-髓质交界区，进一步量化胶原蛋白和弹性蛋白含量、胶原纤维特性和组织内卵泡分布。在培养过程中，原始卵泡的激活伴随着卵巢皮质的松弛，其特征是基质细胞密度从第0天（D0）的3.6±0.2 × 106个细胞/mm3早期下降到第2天（P = 0.033）的2.8±0.1 × 106个细胞/mm3 （P = 0.033）和ECM的动态重塑。值得注意的是，胶原含量从D0时的55.5±1.7%阳性面积逐渐下降到D6时的42.3±1.1% (P = 0.001)，而弹性蛋白含量从D0时的1.1±0.2%上升到D6时的1.9±0.1% （P = 0.001）。纤维连接蛋白和层粘连蛋白含量保持稳定。此外，胶原蛋白和弹性蛋白在整个皮层和培养过程中分布不均匀。亚区水平分析显示，胶原沉积在外皮层最大，中皮层最低（阳性面积分别为69.4±1.2%和53.8±0.8%,P < 0.0001），皮层胶原染色从D0到D2整体下降（65.2±2.4%和60.6±1.8%,P = 0.033），然后趋于稳定。弹性蛋白呈相反分布，最集中在皮层-髓质交界处（3.7±0.6%比0.9±0.2%在外皮层，P < 0.0001），皮层弹性蛋白在D6比D0达到峰值（3.1±0.5%比1.3±0.2%,P < 0.0001）。这一点得到了贯穿皮质的胶原纤维类型的特定特征的证实，表明卵巢皮质ECM的不同表型取决于区域和培养时期，这可能是皮层内观察到的卵泡分布的时空和发育模式的原因。N/A。对剖宫产妇女进行卵巢皮质活检。因此，获得的数据可能不能准确反映非孕妇的ECM分布和结构。明确人类卵巢皮质ECM的组成和结构特征，为进一步探索卵巢微环境奠定基础。这对于理解调节卵泡静止和觉醒的ecm -卵泡相互作用也是至关重要的，从而导致体外激活和体外生长技术的改进。医学研究理事会拨款MR/R003246/1和惠康信托基金会科学合作奖：215625/Z/19/Z。作者没有需要声明的冲突。N/A。
How does in vitro culture alter the human ovarian cortical extracellular matrix (ECM) network structure? The ECM composition and architecture vary in the different layers of the ovarian cortex and are remodelled during in vitro culture. The ovarian ECM is the scaffold within which follicles and stromal cells are organized. Its composition and structural properties constantly evolve to accommodate follicle development and expansion. Tissue preparation for culture of primordial follicles within the native ECM involves mechanical loosening; this induces undefined modifications in the ECM network and alters cell–cell contact, leading to spontaneous follicle activation. Fresh ovarian cortical biopsies were obtained from six women aged 28–38 years (mean ± SD: 32.7 ± 4.1 years) at elective caesarean section. Biopsies were cut into fragments of ∼4 × 1 × 1 mm and cultured for 0, 2, 4, or 6 days (D). Primordial follicle activation, stromal cell density, and ECM-related protein (collagen, elastin, fibronectin, laminin) positive area in the entire cortex were quantified at each time point using histological and immunohistological analysis. Collagen and elastin content, collagen fibre characteristics, and follicle distribution within the tissue were further quantified within each layer of the human ovarian cortex, namely the outer cortex, the mid-cortex, and the cortex–medulla junction regions. Primordial follicle activation occurred concomitantly with a loosening of the ovarian cortex during culture, characterized by an early decrease in stromal cell density from 3.6 ± 0.2 × 106 at day 0 (D0) to 2.8 ± 0.1 × 106 cells/mm3 at D2 (P = 0.033) and a dynamic remodelling of the ECM. Notably, collagen content gradually fell from 55.5 ± 1.7% positive area at D0 to 42.3 ± 1.1% at D6 (P = 0.001), while elastin increased from 1.1 ± 0.2% at D0 to 1.9 ± 0.1% at D6 (P = 0.001). Fibronectin and laminin content remained stable. Moreover, collagen and elastin distribution were uneven throughout the cortex and during culture. Analysis at the sub-region level showed that collagen deposition was maximal in the outer cortex and the lowest in the mid-cortex (69.4 ± 1.2% versus 53.8 ± 0.8% positive area, respectively, P < 0.0001), and cortical collagen staining overall decreased from D0 to D2 (65.2 ± 2.4% versus 60.6 ± 1.8%, P = 0.033) then stabilized. Elastin showed the converse distribution, being most concentrated at the cortex–medulla junction (3.7 ± 0.6% versus 0.9 ± 0.2% in the outer cortex, P < 0.0001), and cortical elastin peaked at D6 compared to D0 (3.1 ± 0.5% versus 1.3 ± 0.2%, P < 0.0001). This was corroborated by a specific signature of the collagen fibre type across the cortex, indicating a distinct phenotype of the ovarian cortical ECM depending on region and culture period that might be responsible for the spatio-temporal and developmental pattern of follicular distribution observed within the cortex. N/A. Ovarian cortical biopsies were obtained from women undergoing caesarean sections. As such, the data obtained may not accurately reflect the ECM distribution and structure of non-pregnant women. Clarifying the composition and architecture signature of the human ovarian cortical ECM provides a foundation for further exploration of ovarian microenvironments. It is also critical for understanding the ECM–follicle interactions regulating follicle quiescence and awakening, leading to improvements in both in vitro activation and in vitro growth techniques. Medical Research Council grant MR/R003246/1 and Wellcome Trust Collaborative Award in Science: 215625/Z/19/Z. The authors have no conflicts to declare. N/A.
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影响因子：
6.7
作者：
Abbara A;Al-Memar M;Phylactou M;Kyriacou C;Eng PC;Nadir R;Izzi-Engbeaya C;Clarke SA;Mills EG;Daniels E;Huo L;Pacuszka E;Yang L;Patel B;Tan T;Bech P;Comninos AN;Fourie H;Kelsey TW;Bourne T;Dhillo WS
通讯作者：
Dhillo WS
CLARITY 揭示了三维卵泡结构和脉管系统的动态
影响因子：
4.6
作者：
Feng Y;Cui P;Lu X;Hsueh B;Möller Billig F;Zarnescu Yanez L;Tomer R;Boerboom D;Carmeliet P;Deisseroth K;Hsueh AJ
通讯作者：
Hsueh AJ
影响因子：
5.9
作者：
Chan CJ;Bevilacqua C;Prevedel R
通讯作者：
Prevedel R
影响因子：
64.8
作者：
Gudipaty SA;Lindblom J;Loftus PD;Redd MJ;Edes K;Davey CF;Krishnegowda V;Rosenblatt J
通讯作者：
Rosenblatt J
DOI：
10.1093/humrep/det388
发表时间：
2014-01
期刊：
Human reproduction (Oxford, England)
影响因子：
--
作者：
Anderson RA;McLaughlin M;Wallace WH;Albertini DF;Telfer EE
通讯作者：
Telfer EE