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. 2013 Sep 2;5(10):1556–1568. doi: 10.1002/emmm.201201584

Figure 4. hNPC treatment in the 5/6 Nx chronic renal injury model.

Figure 4

  • A,B. hNPCs engraft into 5/6 Nx mouse kidneys: (A) hNPCs were labelled with CM-Dil Cell Tracker and injected directly into the renal parenchyma. Mice injected with saline served as controls. Two weeks after cell injection the kidneys were analysed by the maestroCRI in vivo imaging system. Shown are bright field (BF), fluorescence field in red (FF) and merged images of a healthy kidney (naïve, left), remnant kidney treated with hNPCs (middle) or with saline (right). (B) RT-PCR for human-specific β2-microglobulin (hβ2m) demonstrates a representative gel electrophoresis with amplification in the hNPC-injected kidneys, but not in untreated (naïve) or saline-injected kidneys. Mouse β-actin (mβ-actin) served as endogenous control.
  • C,D. hNPCs halt disease progression and improve CrCl: (C) Injection of hNPCs into 5/6 Nx mice led to a significant decrease in the proportion of mice with >15% of renal function deterioration, compared to saline-treated mice. (D) hNPC-treated mice show a significant positive change in CrCl values at 12 weeks when compared to baseline levels (ΔT CrCl) in contrast to saline-treated mice. *p-Value <0.01.
  • E–M. hNPCs integrate into existing tubules and regenerate new tubules: Injected remnant kidneys were stained for the human HLA marker (E–G, J, K—upper panel and M) and a human-specific epithelial marker, panCK/MNF116 (H–L). hNPCs integrated into tubules of host mouse (arrows) and regenerated new tubule-like structures (arrowheads), as demonstrated by IF HLA staining (E). Higher magnification of e is shown in (F and G) HLA staining (green, left) merged with DAPI (overlay, right). (H and I) Various modes of engraftment are highlighted: integration within tubules (H; panCK), integration between tubules (I; panCK) and tubular regeneration (J–M representing consecutive sections). (J) Small early tubules stained for HLA and panCK but not for NCAM1. (K) Mature tubules stained for HLA and panCK. (L) A human panCK expressing tubule (upper image) is further delineated to express the human-specific distal marker, EMA (lower image) (M) Staining for HLA (upper image) and cross-reactive proximal marker ENPEP (lower image) shows human origin of a regenerating proximal tubule type.