Experiment: Biomarker assays to evaluate toxicity and inflammation induced by AAVs in ES cell derived kidney organoids.
PI: Ryuji Morizane, MD, PhD
Description: Human kidney organoids generated from human ES are used to evaluate toxicity and inflammation induced by AAV2, AAV8, and AAV9 that express eGFP under CMV promoter. Organoids are exposed to AAVs at MOI 10^5 . MCP-1 and KIM-1 levels were measured using Luminex (Bioplex 200).
Biomarker Assay:
KIM-1 and MCP-1 levels were measured using the Meso Scale Discovery (MSD) platform
Parent Project: Vascularized kidney organoids on chip for efficacy and toxicity testing of somatic genome editing
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Other experiments in this project: 6
- Evaluation of DNA damage, cellular toxicity and inflammatory markers following saCas9 and gRNAs delivery by AAV2 in kidney organoids.
- AAV tropism in kidney organoids derived from human pluripotent stem cells.
- AAV tropism in kidney organoids cultured under flow
- AAV2/2 transduction efficiencies in kidney organoids cultured under flow determined by FACS
- Biomarker assays to evaluate toxicity induced by AAVs in iPS cell derived kidney organoids.
- Delivery of saCas9 and gRNAs to nephron epithelia by AAV2 in kidney organoids.
Download: Submitted files
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Results |
| Other Measurement | ||||||||
|---|---|---|---|---|---|---|---|---|
| Condition | Sex | Model | Delivery | Vector | Dosage | MCP-1 (pg/mL) | KIM-1 (pg/mL) | Image |
| Control | Female | Human kidney organoid (ESC-derived) | control | |||||
| AAV2/2 | Female | Human kidney organoid (ESC-derived) | AAV | AAV2/2CMVeGFP | MOI=10^5 | |||
| AAV2/8 | Female | Human kidney organoid (ESC-derived) | AAV | AAV2/8CMVeGFP | MOI=10^5 | |||
| AAV2/9 | Female | Human kidney organoid (ESC-derived) | AAV | AAV2/9CMVeGFP | MOI=10^5 | |||
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Biomarker assays to evaluate toxicity and inflammation induced by AAVs in kidney organoids. H9 samples showing MCP-1 and KIM-1 levels using Luminex (Bio-plex 200). The infection rate was kept at 10^5 vg/cell. |
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