Credit: Nayarana Health
Lagasse Lab
McGowan Institute for Regenerative Medicine
Utilizing the Lymph Node as an Ectopic In vivo Bioreactor for Kidney Tissue
Chronic and end-stage kidney diseases require efficient and effective interventions, in which methods from dialysis to tissue transplantation range in these capabilities. However, with the shortage of available donor organs, researchers have needed to pivot towards tissue engineering options.
Credit: Living Bank
Approach
To address this issue, the Lagasse Lab at the University of Pittsburgh is striving to identify the key players and pathways that support kidney organogenesis.
Animal Model
Early-stage, Green Fluorescent Protein (GFP)+ kidney tissue were implanted into adult mouse intraomenta.
Ectopic graft implantation of early stage mouse kidney tissue into adult lymph node and omenta have shown to be promising niches/sites for kidney maturation, indicated by trademarks like glomeruli and tubules.
Credit: Khan Academy
Evaluation Methods
Tissue
Sectioning
Adult mice with the transplanted grafts were sacrificed and their omenta tissue was fixed in OCT for ~5um sectioning using a Cryostat.
Credit: Sakura Europe
Staining & Imaging
Staining of nuclei and biomarkers CD31, Podoplanin, and Synaptopodin were performed to evaluate the presence of specific cell types indicative of glomeruli and tubules growth and maturation. Imaging of each stain and the inherent GFP+ graft were evaluated using a fluorescent microscope.
Nuclei
Hoechst blue dye was used to stain nuclei DNA of all present cell types.
Co-localization of these biomarkers can allow for scientists to predict the presence of cell populations and gain further insight on the development and growth of implanted tissue.
Conclusions
These images contributed to the research done at the Lagasse Lab, seeking to further detail the lymph node and omenta as appropriate niches for kidney tissue maturation within a host. These underlying mechanisms are currently being investigated further
With this experience at the Lagasse Lab, I've gained an appreciation for immunohistochemistry work, which can provide key support to the testing of experimental hypotheses.