带血供淋巴结移植联合治疗性淋巴管生成重建功能性淋巴通路

Obstructive lymphedema is the tissue edema resulted from the disruption of lymphatic functional structure integrity. Although vascularized lymph node transfer has been shown to provide at least some benefit in human lymphedema patients, autologous lymph nodes incorporate into existing lymphatic vasculature at a low frequency. This poor efficiency may compromise the outcome of the operation because connection with lymphatic vessels is required for maintenance and function of the lymph nodes. The goal of the study is to explore the roles and mechanism of transfered lymph node with therapeutic lymphangiogenesis application of stem cells and lymphatic growth factors on reconstruction of lymph drainage function. Initially, we induces adipose derived stem cells (ADSC) to lymphatic endothelial cells (LEC), sequentially, cultures LEC in 3-dimention hyaluronic acid hydrogel with controlled release of VEGF-C theated lymphangiogenesis to produce lymphatic network in vitro, finally transplant them to rat model with lymphatic vessel damage and vascularized lymph node transfer to facilitate the lymphangiogenesis and recanalization of transfered lymph node. The research will provide a new direction in attempt to reconstruct lymphatic drainage function and also an experimental basis for its potencial clinical application.

阻塞性淋巴水肿是由于淋巴系统功能结构的缺损导致淋巴回流障碍而引起的组织水肿。虽然目前带血供淋巴结移植通过引流泵作用引流滞留于组织间的淋巴液，对阻塞性淋巴水肿具有一定的治疗作用，但由于移植淋巴结与受区"淋巴管再生再通效率低"，其存活和引流泵功能的维持具有不确定性的缺点。问题在于移植淋巴结与受区未能建立满足淋巴回流功能的淋巴管通路。本项目探讨应用干细胞移植联合促淋巴管生长因子动员的"治疗性淋巴管生成"方法，提高移植淋巴结与受区淋巴管再生再通效率，实现功能性淋巴通路重建。研究拟应用带血供自体淋巴结移植大鼠模型，通过脂肪干细胞（ADSC）诱导分化为淋巴管内皮细胞（LEC），应用复合VEGF-C的控释透明质酸水凝胶三维培养体外构建初始淋巴管网，再体内植入带血供移植的淋巴结周围出芽增殖成熟，实现淋巴结与受区淋巴管再生再通。本项目为功能性淋巴循环通路重建提供了一个研究方向，并为其临床应用提供实验基础。

本项目研究结果证实小鼠脂肪干细胞（ADSC）和分选的Podolain+亚群细胞在体外能够向淋巴管内皮细胞（LECs）诱导并参与了淋巴管新生。通过优化的体外诱导体系证实小鼠ADSC中Podolain+亚群细胞能够获得效率较高的LECs表型细胞，为淋巴水肿淋巴管新生治疗研究提供了治疗细胞来源。在此基础上应用ADSC来源的Podolain+ 细胞进行细胞移植，证实能够显著促进淋巴管缺损区淋巴管网的形成并缓解淋巴水肿，为阻塞性淋巴水肿治疗提供新的途径。本项目将进一步探讨应用ADSC/Podolain+细胞移植提高移植淋巴结与受区淋巴管再生再通效率的应用途径和机制，实现功能性淋巴通路重建，并为其临床应用提供实验基础。