Systematic Sampling of the Female Reproductive System for Molecular Characterization

Abstract

AbstractAs 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 procurementBasic Protocol 2: Removal of the female reproductive system en blocBasic Protocol 3: Postsurgical dissection of ovariesBasic Protocol 4: Postsurgical dissection of fallopian tubesBasic Protocol 5: Postsurgical dissection of cervixBasic Protocol 6: Postsurgical dissection of uterine bodySupport Protocol 1: OCT‐embedded tissue protocolSupport Protocol 2: Tissue fixation protocolSupport Protocol 3: Snap‐frozen tissue protocolBasic Protocol 7: Tissue slice preparation for Visium analysisSupport Protocol 4: Hematoxylin and eosin staining for 10X Visium imagingBasic Protocol 8: Manual tissue dissociation for Multiome analysisBasic Protocol 9: Tissue dissociation for Multiome analysis using S2 Singulator