A strategy for in vitro propagation of rat nephrons.
BACKGROUND
Recent advances in the understanding of the molecular biology of rodent renal development have lead to the ability to culture the components of the developing rat kidney-the ureteric bud (UB) and the metanephric mesenchyme (MM)-in isolation from one another. Here we here describe a method for subculturing and propagating either whole rat metanephric rudiments or isolated rat UBs. Exploiting the branching program intrinsic to the UB, propagated rat UBs can be recombined with fresh rat mesenchyme to form a large number of rat "neokidneys" derived from a single progenitor that may be amenable to site-specific modulation of function.


METHODS
Whole rat metanephric rudiments or isolated rat UBs were cultured and subdivided through several generations. Both cultured progenitor and subsequent generations of isolated rat UBs were recombined with freshly isolated rat metanephric mesenchyme. The tubules of these rat neokidneys were examined for expression of epithelial markers.


RESULTS
Isolated rat UBs and whole rat metanephric rudiments could be propagated through several generations and appeared morphologically identical to their progenitors. Generations of isolated rat UB could be recombined with fresh rat mesenchyme and the resultant neokidney displayed the same morphologic appearance as the whole rat kidney rudiment. The UB-derived and MM-derived portions of the tubules of these rat neokidneys appear contiguous.


CONCLUSIONS
The recombination of cultured and propagated rat UB with rat mesenchyme yielded rat neokidneys with tubular structures that appeared morphologically identical to whole rat kidney. In vitro propagation of rat metanephric rudiments and recombination of rat UB and MM suggest the possibility of designing nephrons that possess specific desirable functions that can be propagated in vitro.
Steer
Dylan L
DL
Department of Medicine, University of California, San Diego, La Jolla, California 92093, USA.
Bush
Kevin T
KT
Meyer
Tobias N
TN
Schwesinger
Catherine
C
Nigam
Sanjay K
SK
D000818:Animals; D004848:Epithelium / Q000196:embryology; D005260:Female; D016332:Fetal Tissue Transplantation; D008648:Mesoderm; D051379:Mice; D009399:Nephrons / Q000166:cytology / Q000196:embryology* / Q000637:transplantation; D009924:Organ Culture Techniques / Q000379:methods*; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D014183:Transplantation, Heterologous