Jun 24, 2025
Establishment of a mouse model containing blood group antibodies
- lei wang1
- 1Hubei Shizhen Laboratory, Hubei University of Chinese Medicine
- lncyly
Protocol Citation: lei wang 2025. Establishment of a mouse model containing blood group antibodies. protocols.io https://dx.doi.org/10.17504/protocols.io.81wgbkm5ngpk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 24, 2025
Last Modified: June 24, 2025
Protocol Integer ID: 220881
Keywords: abo blood type antigen, local injection of lentivirus, days after the tumor cell line, lentivirus, containing blood group, tumor cell line, abo drug, blood group, mouse armpit, empty virus, mouse model, blood, mouse, antibody, antigen
Funders Acknowledgements:
Scientific researchproject of Hunan Provincial Health Commission
Grant ID: No:202204010038
Natural Science Foundation of Inner Mongolia Autonomous Region
Grant ID: 2021MS08060
Inner Mongolia Medical University Joint Project
Grant ID: YKD2022LH072
Science and Technology Programof the Joint Fund of Scientific Research for the Public Hospitalsof Inner Mongolia Academy of Medical Sciences
Grant ID: 2024GLIH0990
Disclaimer
All pictures used are drawn by the author. No theft or infringement has occurred.
Abstract
Animals were immunized with ABO blood type antigens for the first 3 weeks. Five days after the tumor cell line (2x105/mouse) was inoculated in the mouse armpit, local injection of lentivirus-ABO drugs (1x107/mouse, 200μl) was started, once every 2 days, and the empty virus was used as the control group.
Image Attribution
The datasets used and/or analysed during the current study available from the corresponding author on reasonable request.The pictures are copyrighted by the author.
Guidelines
Chinese Animal Welfare Guidelines（2023）
Article 25 [Welfare Ethics Principles] The use of experimental animals shall follow the principles of substitution, reduction and optimization.
Units and individuals engaged in experimental animal work shall take effective measures to meet the welfare of experimental animals and the ethical requirements of animal experiments.
Article 26 [Welfare Ethics Management] The science and technology department of the State Council shall establish the National Laboratory Animal Welfare Ethics Committee to guide the welfare of experimental animals and animal experiment ethics work across the country.
The science and technology departments of the people's governments of provinces, autonomous regions, and municipalities directly under the Central Government and the Xinjiang Production and Construction Corps shall be responsible for guiding and supervising the welfare and ethical management of experimental animals in their administrative regions.
Article 27 [Welfare Ethics Review] Units engaged in experimental animal work shall establish an experimental animal welfare ethics committee to conduct independent, fair and scientific ethical review and supervision of activities involving experimental animals.
Individuals engaged in experimental animal work shall entrust the welfare ethics committee of similar experimental animal units to conduct reviews when conducting related activities involving experimental animals.
Article 28 [Transport Welfare] When transporting experimental animals, necessary measures shall be taken to meet the welfare requirements of experimental animals during transportation.
Materials
Babl/c mice were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd
200 μL of 5% red blood cell suspension from Shanghai Blood Biomedical Co., Ltd
lentivirus-ABO drugs from Beijing Likeli Biological Company
Establishment of a mouse model containing blood group antibodies
Four-week-old female Babl/c mice were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd. and kept in SPF-grade cages in the Experimental Animal Center of Chifeng Municipal Hospital, with a maximum of 5 mice per cage.
200 μL of 5% red blood cell suspension from Shanghai Blood Biomedical Co., Ltd. was injected intraperitoneally once a week for 3 consecutive weeks.
The mice were killed in the fourth week, and whole blood was collected from the mice by eye removal, and peripheral serum was collected.
Five days after the tumor cell line (2x105/mouse) was inoculated in the mouse armpit, local injection of lentivirus-ABO drugs (1x107/mouse, 200μl) was started, once every 2 days, and the empty virus was used as the control group.
In order to prevent the virus drug from entering the blood circulation and reducing the efficacy, the injection method was selected to be carried out around the tumor and multiple points.
To ensure the welfare of the mice, when the tumor volume reached 1000mm3 or the maximum tumor diameter reached 15mm or tumor ulcers appeared, the mice were immediately killed by cervical dislocation.
The length and width of the mouse tumor were measured and recorded, and the tumor volume was calculated. The specific formula is: V=lengthxwidth2x0.50.
Acknowledgements
We would like to thank the Science and Technology Department of Inner Mongolia, the Health Department of Hunan Province, the Science and Technology Bureau of Chifeng City, and the Chifeng Municipal Hospital for their financial and venue support, and we would like to thank all the patients who provided data information for this study.