Mouse stem cells seeded into decellularized rat kidney scaffolds endothelialize and remodel basement membranes.
BACKGROUND
To address transplant organ shortage, a promising strategy is to decellularize kidneys in a manner that the scaffold retains signals for seeded pluripotent precursor cells to differentiate and recapitulate native structures: matrix-to-cell signaling followed by cell-cell and cell-matrix interactions, thereby remodeling and replacing the original matrix. This would reduce scaffold antigenicity and enable xeno-allografts.


RESULTS
DAPI-labeled cells in arterial vessels and glomeruli were positive for both endothelial lineage markers, BsLB4 and VEGFR2. Rat scaffold's basement membrane demonstrated immunolabeling with anti-mouse laminin β1. Labeling intensified over time with 14 day incubations.


CONCLUSIONS
We provide new evidence for matrix-to-cell signaling in acellular whole organ scaffolds that induces differentiation of pluripotent precursor cells to endothelial lineage. Production of mouse basement membrane supports remodeling of host (rat)-derived scaffolds and thereby warrants further investigation as a promising approach for xenotransplantation.


METHODS
We previously showed that murine embryonic stem cells arterially seeded into acellular rat whole kidney scaffolds multiply and demonstrate morphologic, immunohistochemical and gene expression evidence for differentiation. Vascular cell endothelialization was now further tested by endothelial specific BsLB4 lectin and anti-VEGFR2 (Flk1) antibodies. Remodeling of the matrix basement membranes from rat to mouse ("murinization") was assessed by a monoclonal antibody specific for mouse laminin β1 chain.
Division of Nephrology, Hypertension and Renal Transplantation; University of Florida, Gainesville, FL USA. Rossea@medicine.ufl.edu
Edward A
EA
Ross
Dale R
DR
Abrahamson
Patricia
P
St John
William L
WL
Clapp
Matthew J
MJ
Williams
Naohiro
N
Terada
Takashi
T
Hamazaki
Gary W
GW
Ellison
Christopher D
CD
Batich
D000818:Animals; D001485:Basement Membrane / Q000378:metabolism*; D002454:Cell Differentiation; D019070:Cell Lineage; D024141:Collagen Type IV / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D051379:Mice; D051381:Rats; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D054457:Tissue Scaffolds
D024141:Collagen Type IV; C513277:Lamb1 protein, mouse; D007797:Laminin