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女性生殖系统的系统采样用于分子特征分析。

Systematic Sampling of the Female Reproductive System for Molecular Characterization.

机构信息

Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania.

Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.

出版信息

Curr Protoc. 2023 Aug;3(8):e848. doi: 10.1002/cpz1.848.

DOI:10.1002/cpz1.848
PMID:37584588
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10575691/
Abstract

As part of the National Institutes of Health Human BioMolecular Atlas Program to develop a global platform to map the 37 trillion cells in the adult human body, we are generating a comprehensive molecular characterization of the female reproductive system. Data gathered from multiple single-cell/single-nucleus and spatial molecular assays will be used to build a 3D molecular atlas. Herein, we describe our multistep protocol, beginning with an optimized organ procurement workflow that maintains functional characteristics of the uterus, ovaries, and fallopian tubes by perfusing these organs with preservation solution. We have also developed a structured tissue sampling procedure that retains information on individual-level anatomic, physiologic, and individual diversity of the female reproductive system, toward full exploration of the function and structure of female reproductive cells. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Preparation and preservation of the female reproductive system (ovaries, fallopian tubes, and uterus) prior to procurement Basic Protocol 2: Removal of the female reproductive system en bloc Basic Protocol 3: Postsurgical dissection of ovaries Basic Protocol 4: Postsurgical dissection of fallopian tubes Basic Protocol 5: Postsurgical dissection of cervix Basic Protocol 6: Postsurgical dissection of uterine body Support Protocol 1: OCT-embedded tissue protocol Support Protocol 2: Tissue fixation protocol Support Protocol 3: Snap-frozen tissue protocol Basic Protocol 7: Tissue slice preparation for Visium analysis Support Protocol 4: Hematoxylin and eosin staining for 10X Visium imaging Basic Protocol 8: Manual tissue dissociation for Multiome analysis Basic Protocol 9: Tissue dissociation for Multiome analysis using S2 Singulator.

摘要

作为美国国立卫生研究院人类生物分子图谱计划的一部分，该计划旨在开发一个全球平台，以绘制成年人体 37 万亿个细胞图谱，我们正在对女性生殖系统进行全面的分子特征分析。将使用来自多个单细胞/单细胞核和空间分子分析的收集的数据来构建 3D 分子图谱。在此，我们描述了我们的多步方案，从优化的器官获取工作流程开始，该流程通过向这些器官灌注保存溶液来维持子宫、卵巢和输卵管的功能特性。我们还开发了一种结构化的组织采样程序，该程序保留了女性生殖系统个体水平解剖结构、生理和个体多样性的信息，以充分探索女性生殖细胞的功能和结构。©2023 威立出版公司。基本方案 1：在获取前对女性生殖系统（卵巢、输卵管和子宫）进行准备和保存基本方案 2：整块切除女性生殖系统基本方案 3：卵巢的术后解剖基本方案 4：输卵管的术后解剖基本方案 5：子宫颈的术后解剖基本方案 6：子宫体的术后解剖基本方案 7：用于 Visium 分析的组织切片制备支持方案 1：OCT 包埋组织方案支持方案 2：组织固定方案支持方案 3：冷冻组织方案基本方案 8：用于 Multiome 分析的组织解离基本方案 9：使用 S2 Singulator 进行 Multiome 分析的组织解离

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/08fb/10575691/2253834aefd0/nihms-1917026-f0010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/08fb/10575691/194149197273/nihms-1917026-f0001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/08fb/10575691/2253834aefd0/nihms-1917026-f0010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/08fb/10575691/194149197273/nihms-1917026-f0001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/08fb/10575691/0c09247181de/nihms-1917026-f0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/08fb/10575691/aaf160de63bf/nihms-1917026-f0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/08fb/10575691/f7a5a86ef4b6/nihms-1917026-f0007.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/08fb/10575691/2253834aefd0/nihms-1917026-f0010.jpg

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本文引用的文献

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Endometrial Stem/Progenitor Cells-Their Role in Endometrial Repair and Regeneration.子宫内膜干/祖细胞——它们在子宫内膜修复和再生中的作用
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