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Fig. 1. Rat metanephric mesenchymes recapitulate differentiation of kidney
epithelia in vivo under defined culture conditions. (A) Metanephric
mesenchymes cultured in basal media undergo apoptosis. (B) Addition of
Fgf2 and Tgf
to the culture media induces survival of clusters of
progenitors. (C) These aggregates express Wnt4 as detected by
in situ hybridization. (D) After continued culture with Fgf2 and
Tgf, mesenchymes degenerate without differentiating into epithelia.
(E-J) NHBF, Lif or NGAL, when combined with Fgf2 and Tgf, each
induced continued expansion of metanephric mesenchymes and their
differentiation into organotypic epithelia within 7 days of organ culture.
Tubules stain positive for Cdh1 (E-cadherin; F), whereas glomerular-like
structures express podocalyxin-like (Podxl) (G). Histologically, these
structures resemble kidney epithelia at and beyond the S-shaped body stage
(H-J). (K) The sequence of metanephric mesenchymal differentiation in
organ culture recapitulates epithelial differentiation in vivo (in this case
in the presence of Lif). Arrows delineate a mesenchymal aggregate, after 3-4
days of differentiation in vitro, reminiscent of pretubular aggregates in
vivo. UB, ureteric bud; CM, condensed mesenchyme; PA, pretubular aggregate;
SB, S-shaped body; Tb, tubule; Gl, glomerular-like structure.
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