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.
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 ratUBs. Exploiting the branching program intrinsic to the UB, propagated ratUBs 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 ratUBs were cultured and subdivided through several generations. Both cultured progenitor and subsequent generations of isolated ratUBs were recombined with freshly isolated rat metanephric mesenchyme. The tubules of these rat neokidneys were examined for expression of epithelial markers. RESULTS: Isolated ratUBs and whole rat metanephric rudiments could be propagated through several generations and appeared morphologically identical to their progenitors. Generations of isolated ratUB 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 ratUB 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 ratUB and MM suggest the possibility of designing nephrons that possess specific desirable functions that can be propagated in vitro.
Authors: Mita M Shah; James B Tee; Tobias Meyer; Catherine Meyer-Schwesinger; Yohan Choi; Derina E Sweeney; Thomas F Gallegos; Kohei Johkura; Eran Rosines; Valentina Kouznetsova; David W Rose; Kevin T Bush; Hiroyuki Sakurai; Sanjay K Nigam Journal: Am J Physiol Renal Physiol Date: 2009-09-02
Authors: Eran Rosines; Kohei Johkura; Xing Zhang; Heidi J Schmidt; Marvalyn Decambre; Kevin T Bush; Sanjay K Nigam Journal: Tissue Eng Part A Date: 2010-08 Impact factor: 3.845
Authors: Chakradhar Velagapudi; Rune-Par Nilsson; Myung Ja Lee; Hannah S Burns; Jill M Ricono; Mazen Arar; Veronique L Barnes; Hanna E Abboud; Jeffrey L Barnes Journal: Am J Pathol Date: 2011-12-02 Impact factor: 4.307
Authors: Mita M Shah; Hiroyuki Sakurai; Derina E Sweeney; Thomas F Gallegos; Kevin T Bush; Jeffrey D Esko; Sanjay K Nigam Journal: Dev Biol Date: 2010-01-06 Impact factor: 3.582
Authors: L Perin; S Giuliani; D Jin; S Sedrakyan; G Carraro; R Habibian; D Warburton; A Atala; R E De Filippo Journal: Cell Prolif Date: 2007-12 Impact factor: 6.831