In vivo pulmonary tissue engineering:: Contribution of donor-derived endothelial cells to construct vascularization
In vivo pulmonary tissue engineering:: Contribution of donor-derived endothelial cells to construct vascularization
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DOI：
10.1089/tea.2007.0041
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发表时间：
2008-03-01
影响因子：
4.1
通讯作者：
Finck, Christine M.
中科院分区：
文献类型：
--
作者：
Mondrinos, Mark J.;Koutzaki, Sirma H.;Finck, Christine M.
工程化肺组织肺内植入为儿童和成人肺部疾病的治疗提供了一种潜在的治疗途径。为了实现这一目标，我们报告了利用皮下Matrigel Plug模型在体内生成带血管的肺组织结构。从胚胎17.5天的肺组织中分离出含有上皮细胞、间充质细胞和内皮细胞的小鼠胎肺细胞(FPC)的混合群体。将FPC与Matrigel混合，注射到成年C57/BL6小鼠的前腹壁皮下，以促进体内肺组织构筑的形成。通过将纤维母细胞生长因子2(FGF2)负载的聚乙烯海绵放入水凝胶中，促进血管形成。1周后，尾静脉注射异硫氰酸荧光素标记的葡聚糖后，对供体来源的上皮细胞和内皮细胞进行常规组织学、免疫组织化学染色及血管通畅分析。在仅使用Matrigel的对照组中，检测到一定程度的宿主浸润性，但没有检测到可测量的血管形成。在FPC存在的情况下，这些结构包含导管上皮结构和开放的血管系统。在没有FPC的情况下，外源性FGF2在整个结构中诱导了大量开放的血管的形成；与FPC相结合，它导致了毛细血管密度的增加和发育中的上皮与血管结构之间丰富的界面。这项研究的重要发现是，远端肺上皮细胞分化(通过前表面活性物质蛋白C的表达来评估)可以在体内保持，供体来源的内皮细胞有助于形成与导管上皮结构紧密相连的开放血管。
Intrapulmonary engraftment of engineered lung tissues could provide a potential therapeutic approach for the treatment of pediatric and adult pulmonary diseases. In working toward this goal, we report here on in vivo generation of vascularized pulmonary tissue constructs utilizing the subcutaneous Matrigel plug model. Mixed populations of murine fetal pulmonary cells (FPCs) containing epithelial, mesenchymal, and endothelial cells (ECs) were isolated from the lungs of embryonic day 17.5 fetuses. FPCs were admixed to Matrigel and injected subcutaneously into the anterior abdominal wall of adult C57/BL6 mice to facilitate in vivo pulmonary tissue construct formation. Vascularization was enhanced by placing fibroblast growth factor 2 (FGF2)-loaded polyvinyl sponges into the hydrogel. After 1 week, routine histology and immunohistochemical staining for donor-derived epithelial cells and ECs as well as analysis of patent vasculature in the constructs following tail vein injection of fluorescein isothiocyanate - conjugated dextran were performed. In the Matrigel-only controls, some level of host infiltrate, but no measurable vascularization, was detected. In the presence of FPCs, the constructs contained ductal epithelial structures and patent vasculature. In the absence of FPCs, exogenous FGF2 induced the formation of numerous patent blood vessels throughout the entire constructs; in combination with FPCs, it resulted in enhanced capillary density and abundant interfacing between developing epithelial and vascular structures. The significant findings of this study are that distal pulmonary epithelial differentiation (as assessed by the expression of prosurfactant protein C) can be maintained in vivo and that donor-derived ECs contribute to the formation of patent vessels that interface tightly with ductal epithelial structures.