| In vivo Mouse Mammary Gland Formation
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Author:
Date:
2020-07-05
[Abstract] For years, the mammary gland serves as a perfect example to study the self-renew and differentiation of adult stem cells, and the regulatory mechanisms of these processes as well. To assess the function of given genes and/or other factors on stemness of mammary cells, several In vitro assays were developed, such as mammospheres formation assay, detection of stem cell markers by mRNA expression or flow cytometry and so on. However, the capacity of reconstruction of whole mount in the cleared fat pad of recipient female mice is a golden standard to estimate the stemness of the cells. Here we described a step-by-step protocol for in vivo mammary gland formation assay, including preparation of “cleared” recipients and mammary cells for implantation, the surgery process and ...
[摘要] [摘要 ] 多年来，乳腺一直是研究成体干细胞的自我更新和分化以及这些过程的调控机制的完美例证。为了评估给定的基因和/或其他因素对乳腺细胞的干性，有几个函数体外测定法开发出来，如微球体由mRNA表达形成试验，干细胞标志物的检测或流式细胞术等。然而，在雌性小鼠清除的脂肪垫中整个坐骨的重建能力是估计细胞干性的黄金标准。在这里，我们描述了体内的分步操作方案 乳腺形成测定，包括准备“清除”的受体和用于植入的乳腺细胞，手术过程以及如何评估实验结果。结合通过基因编辑和/或药物处理对乳腺细胞的操作，该方案在乳腺干细胞和乳腺发育的研究中可能非常有用。
[背景 ] 作为哺乳动物最典型的器官之一，乳腺（MG）是外分泌腺，负责泌乳。MG的发育受某些性激素的控制，这些激素的水平精确地调节了MG在不同发育阶段的结构，细胞组成和功能变化（Henigighausen and Robinson，2005）。许多遗传和环境因素都参与了乳腺干细胞的调控和MG的发育。为了研究这些因素的功能和机理，已经开发了几种方法，特别是用于评估乳腺细胞的干性。先前的研究表明，只有MG的基底细胞而非管腔细胞能够在受体雌性小鼠清除的脂肪垫中重建上皮树，这表明乳腺干细胞仅存在于基底谱系中（Van Keymeulen 等，2011）。 ）。后来，包括我们在内的许多研究发现了乳腺干细胞的几种标志物（Prater ...
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| Preparation of Mitotic and Meiotic Metaphase Chromosomes from Young Leaves and Flower Buds of Coccinia grandis
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Author:
Date:
2016-04-05
[Abstract] Somatic chromosomes are usually studied from the root tip cells of the plants for cytological investigations. In dioecious plant species like Coccinia grandis, it is very difficult to get meristematic root tip cells from the mature plants of the respective sex forms. In this report, young leaves of the respective sexual phenotypes were used as tissue samples for mitotic chromosome analysis. For meiotic preparation, flower buds of appropriate size were selected for chromosomal studies. Following protocols could be effectively used for routine chromosome preparations in other plant species as well.
[摘要] 通常从植物的根尖细胞研究体细胞染色体用于细胞学研究。 在雌雄异种植物如Coccinia grandis中，很难从相应性别形式的成熟植物中获得分生组织根尖细胞。 在本报告中，各个性表型的幼叶用作有丝分裂染色体分析的组织样品。 对于减数分裂制备，选择适当大小的花芽用于染色体研究。 以下方案也可以有效地用于其他植物物种中的常规染色体制备。
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| Rice Meiotic Chromosome Spread Preparation of Pollen Mother Cells
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Author:
Date:
2014-07-20
[Abstract] In this protocol, we describe a simple and efficient method for meiotic chromosome spread preparation in rice pollen mother cells. Meiotic chromosome preparation by spreading itself is an important technique for plant cytogenetics (Higgins et al., 2004; Chelysheva et al., 2012; Wang et al., 2009); furthermore, it is a crucial step for applying other cytogenetic methods including Fluorescence in situ hybridization (FISH) and immunostaining.
[摘要] 在这个协议，我们描述一种简单和有效的方法，在水稻花粉母细胞减数分裂染色体传播准备。 通过铺展本身的减数分裂染色体制备是植物细胞遗传学的重要技术（Higgins等人，2004; Chelysheva等人，2012; Wang等人，/em>，2009）; 此外，它是应用其他细胞遗传学方法包括荧光原位杂交（FISH）和免疫染色的关键步骤。
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