Hogan Thea, Yates Andrew, Seddon Benedict
Institute of Immunity and Transplantation, University College London, London, United Kingdom.
Institute of infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom.
Bio Protoc. 2017 Dec 20;7(24):e2650. doi: 10.21769/BioProtoc.2650.
This protocol was developed to generate chimeric mice in which T lymphocytes could be stratified by age on the basis of congenic marker expression. The conditioning drug busulfan is used to ablate host haematopoietic stem cells while leaving the peripheral immune system intact. Busulfan treatment is followed by bone marrow transplantation (BMT), with T-cell depleted donor bone marrow bearing a different congenic marker (CD45.2) to that of the host mouse (CD45.1). New cell production post-BMT can thus be tracked by measuring the fraction of CD45.2 cells over time within a population of interest ( Hogan , 2015 ; Gossel , 2017 ).
本方案旨在培育嵌合小鼠，其中T淋巴细胞可根据同基因标记表达按年龄分层。预处理药物白消安用于清除宿主造血干细胞，同时保持外周免疫系统完整。白消安治疗后进行骨髓移植（BMT），供体骨髓经T细胞去除，带有与宿主小鼠（CD45.1）不同的同基因标记（CD45.2）。因此，通过测量感兴趣群体中CD45.2细胞随时间的比例，可追踪骨髓移植后的新细胞生成情况（霍根，2015；戈塞尔，2017）。
