Microdissection of Primary Renal Tissue Segments and Incorporation with Novel Scaffold-free Construct Technology

Chase A. Arbra; Satish N. Nadig; Sarah Grace Dennis; Sanket Pattanaik; Heather A. Bainbridge; J. Matthew Rhett; Stephen A. Fann; Carl Atkinson; Michael J. Yost

doi:10.3791/57358

A subscription to JoVE is required to view this content. Sign in or start your free trial.

Method Article

Microdissection of Primary Renal Tissue Segments and Incorporation with Novel Scaffold-free Construct Technology

7.1K views

⸱

DOI:

10.3791/57358

⸱

March 27th, 2018

Chase A. Arbra^*¹ , Satish N. Nadig^*¹ , Sarah Grace Dennis¹ , Sanket Pattanaik¹ , Heather A. Bainbridge¹ , J. Matthew Rhett¹ , Stephen A. Fann¹ , Carl Atkinson¹ , Michael J. Yost¹

¹Department of Surgery, Medical University of South Carolina

^* These authors contributed equally

Summary

Tissue-engineered renal constructs provide a solution for the organ shortage and deleterious effects of dialysis. Here, we describe a protocol to micro dissect murine kidneys for isolation of cortico-medullary segments. These segments are implanted into scaffold-free cellular constructs, forming renal organoids.

Abstract

Kidney transplantation is now a mainstream therapy for end-stage renal disease. However, with approximately 96,000 people on the waiting list and only one-fourth of these patients achieving transplantation, there is a dire need for alternatives for those with failing organs. In order to decrease the harmful consequences of dialysis along with the overall healthcare costs it incurs, active investigation is ongoing in search of alternative solutions to organ transplantation. Implantable tissue-engineered renal cellular constructs are one such feasible approach to replacing lost renal functionality. Here, described for the first time, is the microdissection of murine kidneys for isolation of living corticomedullary renal segments. These segments are capable of rapid incorporation within scaffold-free endothelial-fibroblast constructs which may enable rapid connection with host vasculature once implanted. Adult mouse kidneys were procured from living donors, followed by stereoscope microdissection to obtain renal segments 200 - 300 µm in diameter. Multiple renal constructs were fabricated using primary renal segments harvested from only one kidney. This method demonstrates a procedure which could salvage functional renal tissue from organs that would otherwise be discarded.

Introduction

Chronic kidney disease (CKD) is one of the current major public health challenges worldwide¹. The prevalence of CKD in the United States is over 14% of the total population, with over 600,000 Americans suffering from the most severe form, end-stage renal disease (ESRD)². The current treatment options available for those with ESRD include dialysis and kidney transplantation. Although approximately 25,000 patients undergo renal transplantation each year, a significant number of patients are added annually leading to a large disparity between those awaiting a life-saving organ and those receiving transplantation