Oct 11, 2022

Public workspacePost-Surgical Dissection of Uterine Body

DOI
dx.doi.org/10.17504/protocols.io.8epv59yb4g1b/v1
  • 1University of Pennsylvania;
  • 2Gift of Life Donor Program
Stephen Fisher
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DOI: dx.doi.org/10.17504/protocols.io.8epv59yb4g1b/v1
Protocol Citation: 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 2022. Post-Surgical Dissection of Uterine Body. protocols.io https://dx.doi.org/10.17504/protocols.io.8epv59yb4g1b/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 05, 2022
Last Modified: October 11, 2022
Protocol Integer ID: 68277
Funders Acknowledgement:
NIH
Grant ID: U54HD104392
Abstract
This protocol describes dissection of the uterine body in preparation for 10X Visium, 10X Multiomics, pathology review, and biobanking. Various systems can be used to sample and maintain the orientation of biospecimens collected from the uterus. In this protocol we divide the uterus along its true axis and process the anterior and posterior walls of the uterus separately. Other investigators have divided the uterus perpendicular to the true axis which captures both the anterior and posterior walls together at various points along the true axis.
Materials
  • Ice
  • Clean cutting surface
  • Disposable scalpels
  • Kimwipes
  • Disposble rulers (with centimeter measurements)
  • Ice bucket
  • Biohazard bags and container
  • ReagentNuclease-free Water
  • ReagentDMEM/F-12Thermo FisherCatalog #11320033
  • ReagentRPMI 1640 MediumThermo FisherCatalog #11875085
  • ReagentMACS® Tissue Storage SolutionMiltenyi BiotecCatalog #130-100-008
  • ReagentDPBS (10X), no calcium, no magnesium Thermo Fisher ScientificCatalog #14200075 - dilute to 1X with nuclease-free water
  • ReagentMarking dyeCancer DiagnosticsCatalog #03000P
Equipment
VWR® Pour-Boat Weighing Dishes
NAME
flat-bottomed dish
TYPE
VWR International
BRAND
10803-166
SKU



Before start
Dilute phosphate buffered saline (Gibco; 14200-075) to 1X with nuclease-free water.
Ensure you have a clean cutting surface and fresh, disposable surgical blades.
Remove uterus from ice and dry it.
Divide the cervix and uterus into halves in coronal or frontal place.
Identify internal cervical os.
Working on the anterior side of the uterine body, slice it into three horizontal sections from the internal cervical os as the lower limit to the fundus as the upper limit: fundus, anterior uterine body, and anterior lower uterine segment.

Dissection of the Uterine Body. First the coronal plane of the uterus is used to divide the anterior and posterior walls of the uterus. The uterus is then divided into the fundus, uterine body and lower uterine segment (LUS). The LUS is the area of the uterus that undergoes circumferential dilatation during labor. Its inferior anatomic border is the junction of the cervix and uterus (internal cervical os). Here the vesicouterine reflection was used as the superior anatomic border of the LUS. The fundus is the round area at the top of the uterus above the openings of the Fallopian tubes. The remaining area about the LUS and below the fundus is the uterine body.
Tissue layers in the uterus. The outer layer of the uterus, the serosa, consists of a thin layer of epithelial cells. The inner lining of the uterus is the endometrium. The endometrium is comprised primarily of stromal fibroblasts, epithelial cells and immune cells. Proliferation, differentiation and function of the cells in the endometrium varies by the level of reproductive hormones. Then endometrium is the tissue shed in the cyclic process of menstruation. The myometrium is a muscular layer that lies between the serosa and endometrium and is the predominant tissue type in the uterus. In this protocol the anterior or posterior wall of the uterus in each region (lower uterine segment, body and
fundus) are cut in multiple pieces (which vary based on the size of the uterus). Each individual piece is divided into the serosa and myometrium and the endometrium and myometrium which assists with maintenance of orientation.

Remove the anterior lower uterine segment and place the remainder of the specimen on ice.
Divide the anterior lower uterine segment into five sections, from right to left.
Divide each section into four additional sections, from right to left.
Place each of the 20 anterior lower uterine sections on its side and using a razor blade, separate the serosa and myometrium from the submucosal myometrium and endometrium.
Using disposable weigh boats, weigh each of the 40 tissue specimen.
Tissue can be processed with protocols OCT-Embedded Tissue Preparation, Tissue Fixation Preparation, or Snap-Frozen Tissue Preparation depending on the desired downstream processing.
Repeat steps for anterior uterine body, anterior uterine fundus, posterior lower uterine segment, posterior uterine body, posterior uterine fundus.