Failure of bone marrow to reconstitute lung epithelium
Failure of bone marrow to reconstitute lung epithelium
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DOI：
10.1165/rcmb.2005-0175rc
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发表时间：
2005-10-01
影响因子：
6.4
通讯作者：
Mulligan, RC
中科院分区：
文献类型：
--
作者：
Kotton, DN;Fabian, AJ;Mulligan, RC
已经提出了上皮组织重建的新范例，其中来自骨髓的循环细胞有助于形成各种上皮细胞类型。关于肺，最近的几份报告使用免疫荧光显微镜来证明骨髓来源的细胞作为II型肺细胞（肺泡的内源性祖细胞）的植入。我们在这里表明，免疫荧光显微镜，已经在以前的报告中使用，不能可靠地识别罕见的移植细胞在肺组织切片移植后的骨髓细胞或纯化的造血干细胞与无处不在的标签跟踪。我们采用了一个谱系特异性报告系统的基础上，表达GFP报告基因，只在肺上皮细胞（表面活性蛋白C-GFP）的转基因小鼠，以测定移植细胞的流式细胞术，组织学和分子方法。使用这种方法来评估移植受者，包括那些受到博莱霉素incluced肺损伤，我们证明，当自体荧光，死细胞，和污染的血细胞被排除在分析之外，有没有检测到重建肺泡上皮细胞的未分级骨髓细胞或纯化的造血干细胞。
A new paradigm of epithelial tissue reconstitution has been suggested whereby circulating cells derived from bone marrow contribute to a variety of epithelial cell types. With regard to the lung, several recent reports have used immunofluorescence microscopy to demonstrate engraftment of bone marrow-derived cells as type II pneumocytes, the endogenous progenitors of the lung alveolus. We show here that immunofluorescence microscopy, as has been used in previous reports, cannot reliably identify rare engrafted cells in lung tissue sections after transplantation of bone marrow cells or purified hematopoietic stem cells tracked with ubiquitous labels. We have employed a lineage-specific reporter system based on transgenic mice that express the GFP reporter gene only in lung epithelial cells (surfactant protein C-GFP) to assay for engrafted cells by flow cytometry, histology, and molecular methods. Using this approach to evaluate transplant recipients, including those subjected to bleomycin-incluced lung injury, we demonstrate that when autofluorescence, dead cells, and contaminating blood cells are excluded from analysis, there is no detectable reconstitution of lung alveolar epithelial cells by unfractionated bone marrow cells or purified hematopoietic stem cells.