Kidney Week

Abstract: SA-OR49

Simultaneous Generation of Nephron and Renal Stroma via Progenitor Cell Replacement in Animal Fetus

Session Information

• Assessing Disease Risk in Children: A Developmental Perspective
  November 06, 2021 | Location: Simulive, Virtual Only
  Abstract Time: 04:30 PM - 06:00 PM

Category: Development, Stem Cells, and Regenerative Medicine

• 500 Development, Stem Cells, and Regenerative Medicine

Authors

• Saito, Yatsumu, Tokyo Jikeikai Ika Daigaku, Tokyo, Japan

Yatsumu Saito, MD

• Yamanaka, Shuichiro, Tokyo Jikeikai Ika Daigaku, Tokyo, Japan

Shuichiro Yamanaka,

• Matsui, Kenji, Tokyo Jikeikai Ika Daigaku, Tokyo, Japan

Kenji Matsui,

• Matsumoto, Naoto, Tokyo Jikeikai Ika Daigaku, Tokyo, Japan

Naoto Matsumoto,

• Takamura, Tsuyoshi, Tokyo Jikeikai Ika Daigaku, Tokyo, Japan

Tsuyoshi Takamura,

• Fujimoto, Toshinari, Tokyo Jikeikai Ika Daigaku, Tokyo, Japan

Toshinari Fujimoto,

• Tajiri, Susumu, Tokyo Jikeikai Ika Daigaku, Tokyo, Japan

Susumu Tajiri,

• Matsumoto, Kei, Tokyo Jikeikai Ika Daigaku, Tokyo, Japan

Kei Matsumoto,

• Kobayashi, Eiji, Tokyo Jikeikai Ika Daigaku, Tokyo, Japan

Eiji Kobayashi,

• Yokoo, Takashi, Tokyo Jikeikai Ika Daigaku, Tokyo, Japan

Takashi Yokoo,

Background

To solve the organ transplant shortage by regenerative medicine, the whole kidney, including the renal stroma—which plays important roles in homeostasis, such as structural maintenance, hemodynamics, and kidney endocrine function—must be generated. Previously, we successfully generated a rat nephron using mouse kidney as a scaffold by replacing mouse nephron progenitor cells (NPCs) with rat NPCs. Therefore, animal fetuses can potentially generate human kidneys. Herein, we applied progenitor cell replacement to stromal progenitor cells (SPCs) and to NPCs and SPCs to verify the simultaneous generation of renal stroma and nephrons.

Methods

We harvested the metanephroi of green fluorescent protein rats to extract dissociated single cells (DSCs) by enzymatic treatment. SPCs were extracted from these DSCs by cell sorting targeting the platelet-derived growth factor receptor alpha (PDGFRa)-positive fraction. NPCs were extracted by sorting integrin alpha 8-positive fractions from the PDGFRa-negative fraction. We injected the extracted SPC fractions and both NPC and SPC fractions in the nephrogenic zone of the metanephroi of Foxd1-iDTR mice (host SPC removal model) and Six2/Foxd1-iDTR mice (host NPC and SPC removal model), respectively. The metanephroi were organ cultured for 1 week or transplanted into the retroperitoneum of NOD/Shi-scid/IL-2Rγ^null mice and collected after 2 weeks for evaluation with immunofluorescence staining.

Results

In the SPC removal model, mouse SPCs were replaced with rat SPCs in vitro, and rat stroma was extensively generated in mouse kidneys in vivo. Rat SPCs differentiated into various stromal lineage cells, e.g., mesangial cells, interstitial fibroblasts, vascular pericytes, juxtaglomerular cells, and EPO-producing cells. In the two progenitor cell removal models, cap mesenchyme-like structures were formed with aggregated rat NPCs and SPCs around the mouse ureteric bud in vitro. Rat nephrons and renal stroma were generated in the mouse kidney in vivo.

Conclusion

SPC replacement helped generate heterogeneous rat renal stromal lineage cells in the mouse kidney. Simultaneous NPC and SPC replacement enabled the generation of nephrons and renal stroma between different species.