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During the congress, E-Posters will be accessible to all participants on the congress website 24/7, as well as in the E-poster stations in the congress center.
Preparing your E-Poster
Please review the E-Poster format requirements carefully when preparing your E-Poster. Should your E-Poster not meet the mentioned requirements, it may not be displayed as described above.
E-Poster Submission Deadline
Please prepare and upload your E-Poster no later than March 14, 2026 11.59PM CET. After this date, you will no longer be able to prepare and upload your E-poster and it will not be displayed and accessible on the congress website.
Please follow the instructions below to input your abstract title.
Abstract titles should be brief and reflect the content of the abstract.
We aim to generate chimeric kidneys by combining porcine fetal kidneys with human induced pluripotent stem cells (iPSCs). We hypothesize that nephron progenitor cells (NPCs), stromal progenitor cells (SPCs) and ureteric progenitor cells (UPCs) must be of the same species. To create scaffolds for human iPSCs, host lineages of particular porcine cells need selective removal. Although diphtheria toxin receptor (DTR) systems are often used to mediate conditional cell ablation, it is difficult to control and use safely. We developed a safer system; an inducible caspase-9 system (iC9 system), for generating animal scaffolds for human cells. We can replace rat NPCs and SPCs in developing mouse kidney. The next step is to replace UPCs. In order to create a niche for donor UB progenitor transplantation, recipient UPCs must be ablated before transplant donors’ UPCs. Sox9, which is the transcription factor highly expressed in UB tips, is focused on.
When particular cells were ablated, we used an iC9 system. We injected gRNA, Cas9 protein and long ssDNA (EGFP and Sox9) KI vector into B6RGS zygotes. We performed polymerase chain reaction testing to detect the target gene, which was KI vector. This system enabled us to eliminate selective cells easily. We generated Sox9-iC9 mice by using CRISPR-Cas9 to knock in iC9 and a Venus reporter at the Sox9 C-terminus. Heterozygous mice were bred and E13.5-14.5 fetal kidneys were harvested and cultured on transwell inserts. Adding AP20187 (100 nM) and AT406 (50 μM) made those Sox9-positive cells eliminate. Next, dissociated scattered cells (DSCs) from GFP-expressing mice fetal kidneys were transplanted into Sox9-iC9 mice fetal kidneys and cultured on transwell inserts. We also add AT20187 and AT406. After some days, we fixed them and observed tissue.
We could observe Sox9-positive cells eliminate, which is evidenced by reduced Venus fluorescence. The elimination of UB tips prevented NPCs gathering around them. In other words, the number of cap mesenchyme decreased in the group, to which AP20187 and AT406 added. We also checked that no detectable toxicity was observed for AP20187 and AT406. Ablation efficiency exceeded that achieved with a HoxB7-DTR model. Furthermore, when GFP positive cells were transplanted, they increased day by day. The part of them were consisted of renal tubule. Those phenomena were also watched by immunostaining.
We successfully developed Sox9-iC9 mice fetal kidney, whose Sox9 positive cells were eliminated by adding AP20187 and AT406. Furthermore, GFP positive DSCs composed of fetal kidneys can grow in the other kidney after transplantation. When donor’s UPCs connect to recipient’s UPCs and ureter, urine produced by donor kidney cells can excreted via the recipient’s ureter. We are trying to make chimeric kidneys with this system.
