| A Robust Mammary Organoid System to Model Lactation and Involution-like Processes
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Author:
Date:
2021-04-20
[Abstract] The mammary gland is a highly dynamic tissue that changes throughout reproductive life, including growth during puberty and repetitive cycles of pregnancy and involution. Mammary gland tumors represent the most common cancer diagnosed in women worldwide. Studying the regulatory mechanisms of mammary gland development is essential for understanding how dysregulation can lead to breast cancer initiation and progression. Three-dimensional (3D) mammary organoids offer many exciting possibilities for the study of tissue development and breast cancer. In the present protocol derived from Sumbal et al., we describe a straightforward 3D organoid system for the study of lactation and involution ex vivo. We use primary and passaged mouse mammary organoids stimulated with fibroblast growth factor 2 ...
[摘要] [摘要]乳腺是一种高度动态的组织,在整个生殖生活中都会发生变化,包括青春期的生长以及怀孕和进化的重复周期。乳腺肿瘤诊断代表在世界女性最常见的癌症宽。研究的监管机制乳腺的发育是至关重要的理解荷兰国际集团d如何YS调节可导致乳腺癌的发生和发展。三维(3D)乳腺组织体提供了许多令人激动的可能性的研究的组织发育和乳腺癌。在第E存在衍生自协议Sumbal等人,我们描述一个简单的3D类器官系统的研究的泌乳和复古体外。我们使用成纤维细胞生长因子2 (FGF2)和催乳素刺激的原代和传代小鼠乳腺类器官来模拟小鼠乳腺泌乳和内卷过程的三个周期。这种3D模型类器官代表一个有价值的工具来研究后期产后乳腺的发育和乳腺癌,尤其是产后-相关性乳腺癌。
图形摘要:
乳腺类器官的分离和培养程序
[背景技术]的Th e是乳腺的主要功能是提供营养吨经由牛奶产量Ò新生儿。牛逼乳腺他的发展是主要发生在出生后,由几个因素,包括激素和生长因子调控的一个高度动态的过程(Brisken和拉贾拉姆,2006;斯特恩利希特,2006年)。在青春期,激素和生长因子调节基本的胚胎导管树的导管形态发生(Brisken and ...
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| In vitro Differentiation of Human iPSC-derived Cardiovascular Progenitor Cells (iPSC-CVPCs)
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Author:
Date:
2020-09-20
[Abstract] Induced pluripotent stem cell derived cardiovascular progenitor cells (iPSC-CVPCs) provide an unprecedented platform for examining the molecular underpinnings of cardiac development and disease etiology, but also have great potential to play pivotal roles in the future of regenerative medicine and pharmacogenomic studies. Biobanks like iPSCORE ( Stacey et al., 2013; Panopoulos et al., 2017), which contain iPSCs generated from hundreds of genetically and ethnically diverse individuals, are an invaluable resource for conducting these studies. Here, we present an optimized, cost-effective and highly standardized protocol for large-scale derivation of human iPSC-CVPCs using small molecules and purification using metabolic selection. We have successfully applied this protocol ...
[摘要] [摘要 ] 诱导性多能干细胞衍生的心血管祖细胞(iPSC-CVPCs)为检查心脏发育和疾病病因的分子基础提供了前所未有的平台,但在再生医学和药物基因组学的未来中也具有重要作用。像iPSCORE这样的生物库(Stacey 等,2013 ;Panopoulos 等,2017), 其中包含由数百个遗传和种族不同的个体产生的iPSC,是进行这些研究的宝贵资源。在这里,我们为小分子大规模衍生人iPSC-CVPCs和代谢选择纯化提供了一种优化,具有成本效益和高度标准化的方案。我们已经成功地应用了该协议,从154种不同的iPSCORE iPSC品系中获得了iPSC-CVPC,从而获得了大量的高纯度心脏细胞。一个重要的我们的协议的组成部分是Ç ELL Ç onfluency 估计S(ccEstimate ),用于估计当iPSC集单层将达到80%汇合,这是用于发起的iPSC-CVPC推导最佳的时间的自动方法,并且使得协议为易于在具有不同增长率的iPSC系列中使用。此外,我们发现跨iPSC-CVPC的细胞异质性是由于两种截然不同的心脏细胞类型(心肌细胞(CMs)和心外膜衍生细胞(EPDCs))的比例不同导致的,这两种细胞在心脏再生中均具有关键作用。该协议消除了iPSC线到线优化的需要,并且可以轻松地进行调整和扩展,以进行高通量研究或生成大量适用于再生医学应用的细胞。
[背景 ] ...
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