Engineering three-dimensional pulmonary tissue constructs
Engineering three-dimensional pulmonary tissue constructs
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DOI：
10.1089/ten.2006.12.717
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发表时间：
2006-04-01
影响因子：
--
通讯作者：
Finck, Christine M.
中科院分区：
文献类型：
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作者：
Mondrinos, Mark J.;Koutzaki, Sirma;Finck, Christine M.
在本文中，我们报告的工程三维肺组织体外构建。小鼠胚胎第18天胎肺细胞（FPC）的原代分离物由上皮细胞、间充质细胞和内皮细胞的混合群体组成，如通过免疫组织化学和2-D培养物的RT-PCR所评估的。通过检测SpC基因表达和基因产物前表面活性蛋白C的存在，证实了2-D和3-D培养物中肺泡II型（AE 2）细胞的表型。利用Matrigel水凝胶和聚乳酸-共-乙醇酸（PLGA）和聚-L-乳酸（PLLA）的合成聚合物支架分别制成多孔泡沫和纳米纤维基质，产生FPC的三维结构。三维Matrigel构建体含有肺泡形成单位（AFU），所述肺泡形成单位由显示AE 2细胞超微结构的细胞组成，同时表达SpC基因和基因产物。组织特异性生长因子的加入诱导了分支、囊状上皮结构的形成，使人联想到远端肺结构。重要的是，三维培养是诱导表达形态发生相关的远端上皮基因成纤维细胞生长因子受体2（FGFr 2）所必需的。PLGA泡沫和PLLA支架促进FPC的向内生长，如组织学所证明的。然而，尽管存在组织特异性生长因子，这些基质并不支持远端肺上皮细胞的存活。我们的研究结果可能提供了第一步的漫长道路上工程远端肺组织的增强和/或取代功能障碍的天然肺的疾病，如新生儿肺发育不全。
In this paper, we report on engineering 3-D pulmonary tissue constructs in vitro. Primary isolates of murine embryonic day 18 fetal pulmonary cells (FPC) were comprised of a mixed population of epithelial, mesenchymal, and endothelial cells as assessed by immunohistochemistry and RT-PCR of 2-D cultures. The alveolar type II (AE2) cell phenotype in 2-D and 3-D cultures was confirmed by detection of SpC gene expression and presence of the gene product prosurfactant protein C. Three-dimensional constructs of FPC were generated utilizing Matrigel hydrogel and synthetic polymer scaffolds of poly-lactic-co-glycolic acid (PLGA) and poly-L-lactic-acid (PLLA) fabricated into porous foams and nanofibrous matrices, respectively. Three-dimensional Matrigel constructs contained alveolar forming units (AFU) comprised of cells displaying AE2 cellular ultrastructure while expressing the SpC gene and gene product. The addition of tissue-specific growth factors induced formation of branching, sacculated epithelial structures reminiscent of the distal lung architecture. Importantly, 3-D culture was necessary for inducing expression of the morphogenesis-associated distal epithelial gene fibroblast growth factor receptor 2 (FGFr2). PLGA foams and PLLA nanofiber scaffolds facilitated ingrowth of FPC, as evidenced by histology. However, these matrices did not support the survival of distal lung epithelial cells, despite the presence of tissue-specific growth factors. Our results may provide the first step on the long road toward engineering distal pulmonary tissue for augmenting and/or replacing dysfunctional native lung in diseases, such as neonatal pulmonary hypoplasia.