Isolation of Lymphocytes from a Mouse Peyer's Patch

Abstract

Source: Yazicioglu, Y. F., et al. Unraveling Key Players of Humoral Immunity: Advanced and Optimized Lymphocyte Isolation Protocol from Murine Peyer's Patches. J. Vis. Exp. (2018)

In this video, we demonstrate the isolation of lymphocytes from a harvested mouse Peyer's patch. The isolated lymphocytes can be used for further analysis.

Protocol

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

1. Surgical Excision and Tissue Preparation Steps:

1. Surgical removal of the Small Intestine (SI)
   1. Euthanize mice using CO₂ asphyxiation or any equivalent method approved by the institutional animal ethics committee.
   2. Transfer the mouse to a dedicated area for surgical excision. Place the backside down and sanitize the abdomen with 70% ethanol. Perform a laparotomy by cutting the abdominal skin and peritoneum along the midline from the pubi....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
6-well, 12-well, & 96-well plates	Falcon/Corning	353046,353043/3596
50 ml conical tubes	Falcon	3520
40 µm cell strainer	Falcon	352340
10 ml syringe-plunger	Exel INT	26265
RPMI	Corning	15-040-CV
PBS	Corning	21-040-CM
FBS	Atlanta Biologicals	S11150
Orbital shaker	VWR	Model 200
Curved-end scissor
Fine Serrated Forceps
Small curved scissor