Center for Immunology, University of Minnesota Medical School, Minneapolis, Minnesota.
Department of Integrative Biology and Physiology, University of Minnesota Medical School, Minneapolis, Minnesota.
Curr Protoc. 2022 May;2(5):e446. doi: 10.1002/cpz1.446.
Different populations of immune cells rely on their distinct migration patterns for immunosurveillance, immune regulation, tissue specific differentiation, and maturation. It is often important to clarify whether cells are recirculating or tissue resident, or whether tissue-specific cells are derived from blood-borne precursors or a tissue-resident population. Though migration or tissue residency of immune cells critically depends on the expression of different homing molecules (chemokine receptors, tissue retention molecules, etc.), characterization based solely on the expression of homing molecules may not faithfully reflect the migration patterns of immune cells. Therefore, a more reliable method to clarify migration patterns of immune cells is required. Parabiosis is a surgical connection of two mice resulting in a shared circulatory system, which allows reliable distinction of tissue-resident and circulating cells. Here, we describe a set of protocols for parabiosis, including technique details, pitfalls, and suggestions for optimization and troubleshooting. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Preparation of mice for parabiosis surgery Basic Protocol 2: Parabiosis surgery Basic Protocol 3: Recovery and use of mice after parabiosis surgery Basic Protocol 4: Reversal of parabiotic surgery Basic Protocol 5: Analysis of parabionts.
不同群体的免疫细胞依赖其独特的迁移模式来进行免疫监视、免疫调节、组织特异性分化和成熟。通常需要明确细胞是再循环还是组织驻留，或者组织特异性细胞是来自血源性前体还是组织驻留群体。尽管免疫细胞的迁移或组织驻留性取决于不同归巢分子（趋化因子受体、组织保留分子等）的表达，但仅基于归巢分子的表达来进行特征描述可能无法真实反映免疫细胞的迁移模式。因此，需要一种更可靠的方法来阐明免疫细胞的迁移模式。联体共生是将两只小鼠进行手术连接，形成共享的循环系统，从而能够可靠地区分组织驻留细胞和循环细胞。在这里，我们描述了一套联体共生的方案，包括技术细节、注意事项、优化和故障排除建议。© 2022 威立出版社有限公司。 基本方案 1：联体共生手术的小鼠准备 基本方案 2：联体共生手术 基本方案 3：联体共生手术后的小鼠恢复和使用 基本方案 4：联体共生手术的逆转 基本方案 5：联体共生的分析。
