Jan 15, 2025

Public workspaceKAPP-Sen TMC: Kidney donor selection and wedge biopsy preparation

DOI
dx.doi.org/10.17504/protocols.io.j8nlk9nx1v5r/v1
  • 1The Jackson Laboratory for Genomic Medicine, Farmington, CT, USA;
  • 2University of Texas Health Science Center at San Antonio, San Antonio, Texas, United States;
  • 3Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, Boston, MA, USA;
  • 4Department of Genetics and Genome Sciences, University of Connecticut School of Medicine, Farmington, CT, USA;
  • 5Institute for Systems Genomics, University of Connecticut, Farmington, CT, USA;
  • 6Cedars-Sinai Medical Center, Los Angeles, CA, United States
  • Cellular Senescence Network (SenNet) Method Development Community
  • KAPP-Sen TMC
Sergii Domanskyi
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DOI: dx.doi.org/10.17504/protocols.io.j8nlk9nx1v5r/v1
Protocol Citation: Juliana Alcoforado Diniz, Alejandro Peña, Barbara U. Okafor, Elizabeth L. Stoeger, Stefan G. Tullius, Paul Robson, Nicolas Musi 2025. KAPP-Sen TMC: Kidney donor selection and wedge biopsy preparation. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlk9nx1v5r/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: January 10, 2025
Last Modified: January 15, 2025
Protocol Integer ID: 118025
Keywords: KAPP-Sen TMC
Funders Acknowledgements:
National Institute on Aging (NIA) KAPP-Sen Tissue Mapping Collaborative
Grant ID: U54 AG075941
Abstract
This protocol outlines the procedure for obtaining and processing wedge biopsies from donor nephrectomies. After the kidney is removed from the donor, wedge biopsy is taken from the upper pole area in a controlled open approach. These biopsies are preferred over needle biopsies due to lower risk of complications. The samples are initially fixated in 10% neutral buffered formalin (NBF) at room temperature for 24 hours, then transferred to 70% cold ethanol and refrigerated for further paraffin embedding.
Donor selection criteria
Donor selection criteria
Inclusion criteria:
  1. Both sexes
  2. Age > 18 years
  3. All ethnicities
Exclusion criteria:
1. BMI > 32 kg/m2
2. GFR < 60 ml/min/1.73 m2
3. Hypertension with end-organ damage
4. Active malignancy
5. Chronic pulmonary, hepatic, infectious, gastrointestinal, endocrine, metabolic, neurologic, or inflammatory disease
6. Chronic use of systemic steroids
7. Chronic use of anticoagulants
8. Use of agents with senolytic activity
Collection and preservation of the specimen
Collection and preservation of the specimen
Tissue from donor nephrectomy is processed at the back of the operating room, after the kidney has been removed from the donor but before it has been implanted into the recipient.



Note
The study team prefers wedge biopsies compared to needle biopsies. Needle biopsies are usually used in a percutaneous approach, and there is a higher risk of av fistulas linked to this approach. The biopsies are taken as wedge biopsies, usually from the upper pole area.

Safety information
The study team will not take biopsies at the kidney hilum.

A small wedge biopsy is taken after perfusing the kidney on the back table. The wedge biopsy in an open approach (as the kidney is on the back table) that allows for an entirely controlled scenario. The wedge biopsy is usually 1 cm long, 0.4-0.5 cm wide, and 0.1-0.2 cm deep.
Place the wedge biopsy directly in a tube containing 10% neutral buffered formalin (NBF) for fixation.
Option 1, if the specimen is collected during Thursday-Friday:
Store the sample in 10%NBF at room temperature for 24 hours. Transfer sample to cold ethanol 70% and keep in the fridge protected from light. Ship the tube to JAX-GM in cold packs overnight.

Option 2, if the specimen is collected during Monday-Wednesday:
Ship to JAX-GM at room temperature. At JAX-GM, ensuring that the sample was in 10%NBF fixative for at least 24 hours, transfer the sample to cold ethanol 70% and keep in the fridge protected from light until shipment.
Ship the tube in cold packs overnight to JAX-BH for cassette placement and paraffin embedding. The total time in ethanol may vary from 4 to 7 days.