Britton Chance Center and MoE Key Laboratory for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, Hubei 430074, China.
School of Biomedical Engineering, Hainan University, Haikou, Hainan 570228, China.
Theranostics. 2023 Jan 1;13(2):639-658. doi: 10.7150/thno.79953. eCollection 2023.
Hepatic lymphatics are essential for liver homeostasis and immune function. However, the 3D structure and spatial distribution of hepatic lymphatic vessels (LVs) need to be confirmed. Moreover, the molecular information of hepatic lymphatic endothelial cells (LyECs) needs to be further studied. The bottleneck is the lack of specific markers or labeling methods for hepatic lymphatic endothelial cells (LyECs) Here, we proposed a method for the spatiotemporal sequential injection of antibodies (STSI-Ab) to selectively label hepatic LyECs . In addition, we also developed an efficient hepatic LyEC sorting method and performed deep transcriptome sequencing on hepatic LyECs. The STSI-Ab method achieved selective labeling of the mouse hepatic lymphatic network. Three-dimensional fluorescence imaging results of the STSI-Ab mouse liver lobe clearly showed that hepatic LVs entangled with the portal vein but were not present in the central vein. The imaging data inspired a novel hepatic lobule structure model with an added set of LVs in the portal area. Furthermore, deep transcriptome sequencing of isolated hepatic LyECs and Masson's trichrome staining results suggested that hepatic LyECs might be an important source of collagen fibers deposited in the portal area during the process of liver fibrosis and bile duct ligation (BDL). We proposed an STSI-Ab method for selectively labeling hepatic LVs, distinguishing the hepatic LVs from other vessels, and mapping their 3D structure. This study opens an avenue for understanding hepatic lymphatic structure and it will be very beneficial to the study of hepatic LyEC functions.
肝淋巴管对于肝脏稳态和免疫功能至关重要。然而，肝淋巴管（LVs）的 3D 结构和空间分布仍需确认。此外，肝淋巴管内皮细胞（LyECs）的分子信息也需要进一步研究。其瓶颈在于缺乏肝淋巴管内皮细胞（LyECs）的特异性标记物或标记方法。在这里，我们提出了一种时空顺序注射抗体（STSI-Ab）的方法，用于选择性标记肝 LyECs。此外，我们还开发了一种有效的肝 LyEC 分选方法，并对肝 LyECs 进行了深度转录组测序。STSI-Ab 方法实现了对小鼠肝淋巴管网络的选择性标记。STSI-Ab 处理的小鼠肝叶的三维荧光成像结果清楚地表明，肝 LVs 与门静脉缠绕，但不存在于中央静脉中。这些成像数据启发了一种新的肝小叶结构模型，即在门静脉区域增加了一组 LVs。此外，分离的肝 LyECs 的深度转录组测序和 Masson 三色染色结果表明，肝 LyECs 可能是肝纤维化和胆管结扎（BDL）过程中门静脉区域胶原纤维沉积的重要来源。我们提出了一种选择性标记肝 LVs 的 STSI-Ab 方法，可区分肝 LVs 与其他血管，并对其 3D 结构进行成像。这项研究为理解肝淋巴管结构开辟了道路，对研究肝 LyEC 功能将非常有益。
