This video demonstrates the trans-nasal transplantation of induced pluripotent stem cell-derived human microglia (iPSMG) cells into a mouse brain. Hyaluronidase is applied to the nostrils of a microglial-depleted mouse to enhance permeability. iPSMG suspension is administered intranasally, and the mouse is allowed to recover. Growth factor-rich media is administered regularly to support iPSMG survival in the brain tissue.
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Method Article
Transnasal Transplantation of Induced Pluripotent Stem Cell-Derived Human Microglial Cells into the Mouse Brain
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August 7th, 2025
In This Article
Abstract
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Preparation of cell medium, transplantation medium, and anesthesia mixture
- Prepare the cell medium by adding 10% fetal bovine serum and 0.1% penicillin/streptomycin in Dulbecco's modified Eagle medium (DMEM).
- Prepare the transplantation medium by adding human colony stimulating factor 1 or hCSF1 (250 ng/mL) and human transforming growth factor beta 1 or hTGF-β1 (100 ng/mL) to the cell medium.
- Prepare the anesthesia mixture for intraperitoneal injection by mixing 0.45 mL of medetomidine hy....
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Dulbecco's Modified Eagle Medium (DMEM) | Thermo Fisher Scientific | 10566 | |
| AIN 93G | Oriental Yeast Co | ||
| Fetal bovine serum | GE Healthcare Life Sciences | SH30070.03 | |
| Frozen iPSMG | Shionogi & Co., Ltd | Laboratory for Drug Discovery and Disease Research | |
| Human colony stimulating factor 1 (hCSF1) | PeproTech | 300-25 | |
| Hyaluronidase | Sigma-Aldrich | H-3506 | |
| Medetomidine hydrochloride | Meiji Seika | VETLI5 | |
| Midazolam | Astellas | 18005A2 | |
| Paraformaldehyde | Wako Pure Chemical Industries | 162-16065 | |
| Penicillin/streptomycin | Thermo Fisher Scientific | 15140-122 | |
| Pipette | Eppendorf | 3120000011 | |
| Pipette tip | Eppendorf | 30076028 | |
| PLX5622 | Amadis Chemical | A930097 | |
| Transforming growth factor-β1 (Tgf-b1) | PeproTech | 100-21 |
