Identification of UACA, EXOSC9, and TMX2 in bovine periosteal cells by mass spectrometry and immunohistochemistry
Identification of UACA, EXOSC9, and TMX2 in bovine periosteal cells by mass spectrometry and immunohistochemistry
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DOI：
10.1007/s00216-014-7673-3
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发表时间：
2014-09-01
影响因子：
4.3
通讯作者：
Akiyama, Mari
中科院分区：
文献类型：
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作者：
Akiyama, Mari
检查用于移植的患者来源的细胞是非侵入性的。因此，使用移植前培养的细胞的上清液进行蛋白质组学分析是有益的。为了研究牛骨膜细胞的细胞生态位，这些培养的细胞的上清液进行2-D电泳，然后通过质谱，鉴定1型胶原和3型胶原的C-末端。仅在上清液中发现来自3型胶原的C-末端肽。已知3型胶原可以在细胞内或细胞外表达。接下来通过免疫组织化学检查培养细胞的石蜡切片，其揭示除了C-末端肽之外的3型胶原区域存在于牛骨膜细胞周围，但在上清液中未发现。全长3型胶原与细胞密切相关，只有C-末端肽在培养上清液中可检测到。质谱分析的部分肽数据结合免疫组化也表明，葡萄膜自身抗原与卷曲螺旋结构域和锚蛋白重复序列（UACA），外泌体复合物组分RRP 45（EXOSC 9），和硫氧还蛋白相关的跨膜蛋白2（TMX 2）在牛骨膜细胞中表达。本研究的结果表明，细胞移植前培养上清液的分析可以提供有用的生物标志物，指示用于移植的细胞的生态位。
Inspection of patient-derived cells used in transplantation is non-invasive. Therefore, proteomics analysis using supernatants of cells cultured before transplantation is informative. In order to investigate the cell niche of bovine periosteal cells, supernatants of these cultured cells were subjected to 2-D electrophoresis followed by mass spectrometry, which identified type 1 collagen and the C-terminus of type 3 collagen. Only the C-terminal peptide from type 3 collagen was found in supernatants. It is known that type 3 collagen may be expressed intra- or extra-cellularly. Paraffin sections of the cultured cells were next examined by immunohistochemistry, which revealed that type 3 collagen regions besides the C-terminal peptide were present around the bovine periosteal cells but were not found in supernatants. Full-length type 3 collagen was closely associated with the cells, and only the C-terminal peptide was detectable in culture supernatants. Mass spectrometry analysis of partial peptide data combined with immunohistochemistry also indicated that uveal autoantigen with coiled coil domains and ankyrin repeats (UACA), exosome complex component RRP45 (EXOSC9), and thioredoxin-related transmembrane protein 2 (TMX2) were expressed in bovine periosteal cells. Results of this study indicate that analysis of culture supernatants before cell transplantation can provide useful biomarkers indicating the niche of cells used for transplantation.