Systematic Sampling of the Female Reproductive System for Molecular Characterization

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