Transplantation of Human-Derived Interneuron Precursor Cells into the Mouse Pup Hippocampus

Abstract

Source: Gonzalez-Ramos, A., et. al., Transplantation of Human Stem Cell-Derived GABAergic Neurons into the Early Postnatal Mouse Hippocampus to Mitigate Neurodevelopmental Disorders. J. Vis. Exp. (2022)

This video demonstrates the stereotaxic injection of human-derived interneuron precursor cells into the hippocampus of a genetically modified mouse pup with reduced inhibitory interneurons, aiming to restore excitatory-inhibitory balance.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Intrahippocampal cell transplantation

NOTE: All the steps in this section are performed outside the cell culture hood in the animal facility. Early postnatal transplantation of cells into the brain was performed on P2, considering P0 on the day of birth.

1. Prepare for the transplantation experiment by following the steps below.
   1. Autoclave all the surgical material or, if not possible, sterilize with another approved method.
   2. Mo....

Results

Figure 1: Stereotaxic transplantation in newborn mouse pups at P2. (

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
30 G needle	B Braun	4656300
33 G needle for Hamilton syringe	Hamilton	7762-06
B6.129(Cg)-Cntnap2tm1Pele/J	Jackson Laboratory	17482	Animal model
C57BL/6J	Janvier Labs		Animal model
Digital Stereotax	KOPF	Model 940
Doxycyclin	Sigma-Aldrich	D9891	2 µg/mL
Ethanol	Solveco		70%, 95%, 99.8%
H1 (WA01) ESC	WiCell	WA01	Human embryonic stem cell line under a MTA agreement
Hamilton Syringe	Hamilton	7634-01	5 µL
Isoflurane Baxter	Apoteket AB
Mouse adaptor to stereotax	RWD	68030
Puromycin (Dihydrochloride)	Gibco	A1113803
Tweezer	VWR