Induction of Graft Versus Host Disease in Mouse Models via Allogeneic Bone Marrow and Splenic Cell Transplantation

Published: October 31, 2023

Abstract

Source: Buchele, V., et al. Induction of Intestinal Graft-versus-host Disease and Its Mini-endoscopic Assessment in Live Mice. J. Vis. Exp. (2019).

This video demonstrates the induction of intestinal graft-versus-host disease, GvHD, in a lethally-irradiated recipient mouse using allogeneic hematopoietic stem cells and GvHD-inducing splenic alloreactive T lymphocytes. This protocol helps establish a severe acute intestinal GvHD model to understand the mechanism of the disease and develop innovative therapeutic options.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. GvHD induction Day 0: Total body irradiation of the recipient mice Use female CD45.2+ H2kd+ BALB/c mice that are at least 10 weeks old as recipients. Weigh and record the weight of the recipient mice prior to GvHD induction. NOTE: Ensure t…

Divulgations

The authors have nothing to disclose.

Materials

BIOBEAM 2000 Gamma Irradiator Gamma-Service Meical GmbH
Phosphate-buffered saline (PBS ) Sigma-Aldrich Co. LLC. D8662-6x500ML
Semken Forceps (lenght: 13 cm; serrated, curved tips) Fine Science Tools 11009-13
Hardened Fine Scissors (lenght: 8,5 cm; straight tips; cutting edge: 24 mm) Fine Science Tools 14090-09
RPMI-1640 Medium Sigma-Aldrich Co. LLC. R8758-500ML
Hypodermic needle (26G) B. Braun Melsungen AG 4657683
1 mL syringe B. Braun Melsungen AG 9166017V
50 mL tube Sarstedt Ag & Co.KG 6,25,47,254
Cell strainer with a 40 µm mesh screen BD Falcon 352340
Insulin syringe with sterile interior (30G) BD 324826
Pan T Cell Isolation Kit II, mouse Miltenyi Biotec GmbH 130-095-130 magnet cell separation to isolate splenic T cells
CD90.2 MicroBeads, mouse Miltenyi Biotec GmbH 130-049-101 magnet cell separation to isolate T cell-depleted bone marrow cells
Biopsy Forceps KARL STORZ SE &Co KG 61071 ZJ part of the experimental setup for colonoscopy
Ethylenediaminetetraacetic acid disodium salt dihydrate (Na2EDTA) Carl Roth GmbH & Co.KG 8043.2 ingredient of ACK lysing buffer
Alexa Fluor 700 anti-mouse CD45.2 Antibody (clone: 104; lot: B252126; RRID: AB_493731) Biolegend 109822
PE/Cy7 anti-mouse CD45.1 Antibody (clone: A20; lot: B217246; RRID: AB_1134168) Biolegend 110730
APC/Cy7 anti-mouse CD8a Antibody (clone: 53-6.7, lot: B247008; RRID: AB_312753) Biolegend 100714
Forane / Isofluran AbbVie Inc. B506
Filtrated bovine serum Pan Biotec P40-47500 ingredient of FACS buffer

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Citer Cet Article
Induction of Graft Versus Host Disease in Mouse Models via Allogeneic Bone Marrow and Splenic Cell Transplantation. J. Vis. Exp. (Pending Publication), e21718, doi: (2023).

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