Techniques of fragile renal organoids transplantation in mice

Matsumoto, Naoto; Matsui, Kenji; Saito, Yatsumu; Takamura, Tsuyoshi; Yamanaka, Shuichiro; Yokoo, Takashi; Kobayashi, Eiji

doi:10.1590/acb361102

Cited by 6 publications

(6 citation statements)

References 17 publications

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“…In preclinical xenotransplantation of adult kidneys from pigs to monkeys, we have shown that orthotopic transplantation is superior ( 24 ). For the transplantation of renal organoids, we have previously developed a simple method of transplanting them under the renal membrane without damage ( 10 ). In the present study, we transplanted them under the renal capsule of the host and could generate mature renal tissues between different species.…”

Section: Discussionmentioning

confidence: 99%

“…Single cells from mouse and rat fetal kidneys were obtained following a method ( 10 ) that was a partial modification of a method reported previously ( 11 ). In brief, pregnant E13.5 B6 mice or E15.5 GFP rats were anesthetized via the inhalation of isoflurane (2817774, Pfizer, New York, NY, USA).…”

Section: Methodsmentioning

confidence: 99%

“…Renal organoids were transplanted under the renal capsule using a previously described method ( 10 ). The following is a brief description of the procedure.…”

Section: Methodsmentioning

confidence: 99%

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Beneficial Impact of Interspecies Chimeric Renal Organoids Against a Xenogeneic Immune Response

et al. 2022

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BackgroundAnimal fetal kidneys have the potential to be used as scaffolds for organ regeneration. We generated interspecies chimeric renal organoids by adding heterologous rat renal progenitor cells to single cells from mouse fetal kidneys and applying the renal development mechanism of mouse fetuses to rat renal progenitor cells to examine whether rat renal progenitor cells can differentiate into renal tissues of the three progenitor cell lineages of kidneys between different species. Furthermore, we investigated whether chimeric renal organoids with an increased proportion of recipient cells reduce xenogeneic rejection.MethodsC57BL/6JJmsSlc mice (B6 mice) and Sprague-Dawley-Tg (CAG-EGFP) rat (GFP rats) fetuses were used as donors, and mature male NOD/Shi-scid, IL-2RγKO Jic mice (NOG mice) and Sprague-Dawley rats (SD rats) were used as recipients. First, fetal kidneys were removed from E13.5 B6 mice or E15.5 GFP rats and enzymatically dissociated into single cells. These cells were then mixed in equal proportions to produce chimeric renal organoids in vitro. The chimeric organoids were transplanted under the renal capsule of NOG mice, and maturation of the renal tissues in the organoids was observed histologically. Furthermore, chimeric organoids were prepared by changing the ratio of cells derived from mouse and rat fetal kidneys and transplanted under the renal capsule of SD rats subjected to mild immunosuppression to pathologically analyze the strength of the xenogeneic immune response.ResultsThe cap mesenchyme was reconstructed in vitro, and nephron progenitor cells and ureteric buds were mosaically comprised GFP-negative mouse and GFP-positive rat cells. In the in vivo environment of immunodeficient mice, chimeric renal organoids mosaically differentiated and matured into renal tissues of three lineages. Chimeric renal organoids with high rates of recipient rat cells showed milder rejection than complete xenograft organoids. The vessels of recipient rats entered from the periphery of the transplanted chimeric renal organoids, which might reduce their immunogenicity.ConclusionInterspecies chimeric renal organoids may differentiate into mature renal tissues of each renal progenitor cell lineage. Furthermore, they may reduce transplant rejection compared with xenograft organoids.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Beneficial Impact of Interspecies Chimeric Renal Organoids Against a Xenogeneic Immune Response

et al. 2022

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“…We used our previous method [ 18 , 19 ] that was based on a previous report [ 20 ]. Specifically, E14.5 GFP mice were euthanized by cervical dislocation, and their abdomens were opened to collect embryos.…”

Section: Methodsmentioning

confidence: 99%

Development of a Cryopreservation Technique for Xenogeneic Kidney Grafts: Evaluation Using a Mouse Model

Takamura

Nagashima

Matsunari

et al. 2022

JCM

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To align the xeno-metanephros and renal progenitor cell timing for transplantation treatments, cryopreservation techniques and an efficient transportation of regenerated renal products such as xeno-metanephroi and renal progenitor cells should be established. Therefore, we propose a novel method of xenogeneic regenerative medicine for patients with chronic kidney disease by grafting porcine fetal kidneys injected with human renal progenitor cells. To develop a useful cryopreserve system of porcine fetal kidney and human renal progenitor cells, we examined the cryopreservation of a fetal kidney implanted with renal progenitor cells in a mouse model. First, we developed a new method for direct cell injection under the capsule of the metanephros using gelatin as a support for unzipped fetal kidneys. Then, we confirmed in vitro that the nephrons derived from the transplanted cells were regenerated even after cryopreservation before and after cell transplantation. Furthermore, the cryopreserved chimeric metanephroi grew, and regenerated nephrons were observed in NOD. We confirmed that even in cryopreserved chimeric metanephroi, transplanted cell-derived nephrons as well as fresh transplants grew.

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“…Single cells from mouse and pig fetal kidneys were obtained following a previously reported method [20,25,44]. Fetal kidneys were harvested from the decapitated fetuses under an operating microscope and collected in 1.5 mL tubes containing a minimum essential medium (MEMa; Thermo Fisher Scientific, Inc.).…”

Section: Single Cell Extraction From Fetal Kidneysmentioning

confidence: 99%

Generation of human-pig chimeric renal organoids using iPSC technology

Fujimori,

Yamanaka,

Shimada

et al. 2024

Preprint

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The potential of using porcine organs and human induced pluripotent stem cell (iPSC)-derived organoids as alternative organs for human transplantation has garnered growing attention. However, both approaches still face technological challenges. Interspecies chimeric organ production using human iPSCs is expected to be another promising approach that addresses the challenges associated with organ production. Our research group successfully generated human-mouse chimeric renal organoids by utilizing human iPSC-derived nephron progenitor cells (NPCs) and fetal mouse kidneys. However, the current technology has limited engraftment and development capabilities for human NPCs, and there have been no reports of generating interspecies chimeric renal organoids in larger animals, limited only to rodents. Therefore, in this study, we embarked on the production of human-pig chimeric renal organoids using the pig kidney, which is considered the most promising source of organs for interspecies transplantation to humans. To construct a human-pig chimeric renal organoid culture system, we first modified the existing human-mouse chimeric renal organoid culture system and developed a method that enables the survival and continued renal development of both species. This method was found to be applicable to porcine fetal kidney cells, and ultimately, we successfully produced human-pig chimeric renal organoids. Furthermore, this culture method can also be applied to the generation of human interspecies chimeric kidneys for future clinical applications. The findings of this study serve as a foundational technology that will greatly accelerate future research in humanized pig kidney production for clinical purposes, and are also expected to be used as an evaluation technique to ensure the quality of human NPCs for xenotransplantation.

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Techniques of fragile renal organoids transplantation in mice

Cited by 6 publications

References 17 publications

Beneficial Impact of Interspecies Chimeric Renal Organoids Against a Xenogeneic Immune Response

Beneficial Impact of Interspecies Chimeric Renal Organoids Against a Xenogeneic Immune Response

Development of a Cryopreservation Technique for Xenogeneic Kidney Grafts: Evaluation Using a Mouse Model

Generation of human-pig chimeric renal organoids using iPSC technology

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