Intraspinal Transplantation of Mouse and Human Neural Precursor Cells
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- Abstract
- Table of Contents
- Materials
- Figures
- Literature Cited
Abstract
This unit describes the preparation and transplantation of human neural precursor cells (hNPCs) and mouse neural precursor cells (mNPCs) into the thoracic region of the mouse spinal cord. The techniques in this unit also describe how to prepare the mouse for surgery by performing a laminectomy to expose the spinal cord for transplantation. NPCs genetically labeled with eGFP transplanted into the spinal cord of a mouse following viral?mediated demyelination can efficiently be detected via eGFP expression. Transplantation of these cells into the spinal cord is an efficacious way to determine their effects in neurological disorders such as multiple sclerosis, Alzheimer's disease, and spinal cord injury. Curr. Protoc. Stem Cell Biol . 26:2D.16.1?2D.16.16. © 2013 by John Wiley & Sons, Inc.
Keywords: human neural precursor cells; mouse neural precursor cells; laminectomy; intraspinal transplantation; multipotency
Table of Contents
- Introduction
- Basic Protocol 1: Preparation of hNPCs for Mouse Intraspinal Transplantation
- Support Protocol 1: Verifying NPC Phenotype of hNPCs
- Alternate Protocol 1: Preparation of mNPCs for Intraspinal Transplantation
- Basic Protocol 2: Preparation of Mice for Intraspinal Transplantation
- Basic Protocol 3: Intraspinal Transplantation of mNPCs or hNPCs and Post‐Operative Care
- Reagents and Solutions
- Commentary
- Literature Cited
- Figures
Materials
Basic Protocol 1: Preparation of hNPCs for Mouse Intraspinal Transplantation
Materials
Support Protocol 1: Verifying NPC Phenotype of hNPCs
Materials
Alternate Protocol 1: Preparation of mNPCs for Intraspinal Transplantation
Materials
Basic Protocol 2: Preparation of Mice for Intraspinal Transplantation
Materials
Basic Protocol 3: Intraspinal Transplantation of mNPCs or hNPCs and Post‐Operative Care
Materials
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Figures
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Figure 2.D1.1 Fluorescent microscopy of hNPCs at time of dissociation for preparation for intraspinal transplantation. PAX6 and Nestin are markers for multipotent neural precursor cells and Dapi stains cell nuclei. Images are shown at 40× magnification. View Image -
Figure 2.D1.2 (A ) Setup of stereotaxic apparatus with universal holder with needle support foot on right and electrode holder with removable open side clamp on left of dual small animal stereotaxic platform and tri‐fold paper towels. (B ) The image shows the standard settings for the stereotaxic apparatus to align the needle with the spinal cord at a 70° angle. View Image -
Figure 2.D1.3 Coronal section of a C57Bl/6 mouse transplanted intraspinally with GFP‐mNPCs. Mouse was sacrificed at day 21 post‐transplant, 4% paraformaldehyde perfused, and 10‐µm coronal sections were cut. Transplanted GFP‐mNPCs survived and migrated into the white matter. Engrafted GFP‐mNPCs are green and Dapi‐stained cell nuclei are blue. View Image
Videos
Literature Cited
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