Abstract: SA-PO548

Generation of Interspecies Chimeric Nephrons from Nephron Progenitor Cells by Conditional Elimination and Replacement

Session Information

  • Developmental Biology
    November 04, 2017 | Location: Hall H, Morial Convention Center
    Abstract Time: 10:00 AM - 10:00 AM

Category: Developmental Biology and Inherited Kidney Diseases

  • 401 Developmental Biology

Authors

  • Yamanaka, Shuichiro, Jikei University School of Medicine, Minato, Tokyo, Japan
  • Matsumoto, Kei, Jikei University School of Medicine, Minato, Tokyo, Japan
  • Tajiri, Susumu, Jikei University School of Medicine, Minato, Tokyo, Japan
  • Fujimoto, Toshinari, Jikei University School of Medicine, Minato, Tokyo, Japan
  • Fukunaga, Shohei, Jikei University School of Medicine, Minato, Tokyo, Japan
  • Yokoo, Takashi, Jikei University School of Medicine, Minato, Tokyo, Japan
Background

The animal embryonic organ niche may have applications in the generation of regenerative organs, and progenitor cells may be an appropriate cell source for this purpose because of their safety and availability. Therefore, we established a combination method through which donor cells could be precisely injected into the nephrogenic zone and native nephron progenitor cells (NPCs) be eliminated in a time- and tissue-specific manner. We successfully removed Six2+ NPCs within the nephrogenic niche and replaced transplanted NPCs. The transplanted cells differentiated into neo-nephrons. Furthermore, we generated rat neo-nephrons in the mouse nephrogenic zone by conditional elimination and replacement.

Methods

We used Cre-LoxP technology in combination with diphtheria toxin receptor-loxP (DTR-loxP) and Six2-Cre mice for Diphtheria Toxin (DT)-mediated NPC elimination. Metanephros (MN) was isolated from the Six2/DTR +/+ mouse embryos. Wild-type rat NPCs were transplanted into the Six2/DTR +/+ mouse nephrogenic zone and simultaneously administered DT, followed by co-culture in an organ culture dish for seven days. We examined donor NPCs differentiation into neo nephrons by immunostaining of nephron markers (WT1, PAX8, GATA3, Cytokeratin8, E-cadherin, LTL).

Results

Donor rat NPCs were observed in the broad engraftment in host mouse cap mesenchyme, which eliminated native mouse NPCs on DT administration. The interspecies chimeric nephrons expressed glomerular and tubular markers. We also observed a connection between host collecting ducts and the neo-nephrons.

Conclusion

Using progenitor cells conditional elimination system, we demonstrated that donor rat NPCs replaced host mouse NPCs in the mouse nephrogenic zone, and that generation of neo-nephrons is possible by other species. Thus, this technique enables the differentiation of progenitor cells into nephrons, providing insight into the nephrogenesis and organ regeneration processes. We believe that this technique could effectively be used to evaluate the differentiation of NPCs from pluripotent stem cells (PSCs).

Funding

  • Other NIH Support