Protocols for Molecular Characterization of the Female Reproductive System
Stephen Fisher, Marielena Grijalva, Rong Guo, Sarah A Johnston, Hieu Nguyen, John Renz, Jean G Rosario, Steven Rudich, Brian Gregory, Junhyong Kim, Kate O'Neill
Abstract
Here we describe our multi-step protocol to generate a comprehensive molecular characterization of the female reproductive system. We begin with organ procurement that maintains functional characteristics of the uterus, ovaries, and Fallopian tubes and then describe our structured tissue sampling procedure that represents anatomical, physiological, and individual diversity of the female reproductive system, toward full exploration of the function and structure of female reproductive cells.
These protocols were developed as part of the National Institutes of Health (NIH) Human BioMolecular Atlas Program (HuBMAP), we are centered around the 10X Genomics Multiome and 10X Genomics Visium platforms.
Before start
Steps
Preparation and Preservation of the Female Reproductive System (ovaries, Fallopian tubes and uterus) Prior to Procurement
Removal of the Female Reproductive System En Bloc
Post-Surgical Dissection of Ovaries
Post-Surgical Dissection of Fallopian Tubes
Post-Surgical Dissection of Cervix
Post-Surgical Dissection of Uterine Body
Tissue Slice Preparation for Visium Analysis
Manual Tissue Dissociation for Multiome Analysis
Tissue Dissociation for Multiome Analysis Using S2 Singulator
