Utility of human placental alkaline phosphatase as a genetic marker for cell tracking in bone and cartilage
Utility of human placental alkaline phosphatase as a genetic marker for cell tracking in bone and cartilage
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DOI：
10.1007/s00418-007-0286-6
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发表时间：
2007-06-01
影响因子：
2.3
通讯作者：
Erben, Reinhold G.
中科院分区：
文献类型：
--
作者：
Unger, Nina J.;Odoerfer, Kathrin I.;Erben, Reinhold G.
本研究的目的是评价人胎盘碱性磷酸酶（hPLAP）作为骨和软骨中细胞追踪的遗传标记的实用性，使用在普遍存在的R26启动子[F344-Tg（R26-hPLAP）]控制下表达hPLAP的转基因Fischer 344大鼠。hPLAP酶活性在石蜡和甲基丙烯酸甲酯（MMA）包埋期间保留，并且使用40%乙醇作为固定剂保存得最好。内源性碱性磷酸酶活性可以完全阻断热灭活石蜡和MMA切片，允许在完全没有背景染色的情况下进行hPLAP的组织化学检测。此外，使用免疫组织化学也可以灵敏地检测hPLAP。F344-Tg（R26-hPLAP）大鼠在造血骨髓细胞和基质细胞如成骨细胞、骨细胞和软骨细胞中表现出hPLAP的普遍表达。破骨细胞仅弱表达hPLAP。总之，hPLAP在石蜡和塑料切片中提供了极好的检测质量，并且构成了用于硬组织和软组织中细胞追踪的优异遗传标记。
It was the aim of the current study to evaluate the utility of human placental alkaline phosphatase (hPLAP) as a genetic marker for cell tracking in bone and cartilage, using transgenic Fischer 344 rats expressing hPLAP under the control of the ubiquitous R26 promoter [F344-Tg(R26-hPLAP)]. hPLAP enzyme activity was retained during paraffin and methylmethacrylate (MMA) embedding, and was best preserved using 40% ethanol as fixative. Endogenous alkaline phosphatase activity could be completely blocked by heat inactivation in paraffin and MMA sections, allowing histochemical detection of hPLAP in the complete absence of background staining. In addition, sensitive detection of hPLAP was also possible using immunohistochemistry. F344-Tg(R26-hPLAP) rats demonstrated ubiquitous expression of hPLAP in hematopoietic bone marrow cells and stromal cells such as osteoblasts, osteocytes, and chondrocytes. Osteoclasts only weakly expressed hPLAP. In conclusion, hPLAP provides superb detection quality in paraffin and plastic sections, and constitutes an excellent genetic marker for cell tracking in hard and soft tissues.