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目的探索成人甲状旁腺细胞培养方法的可行性。方法采用胶原酶消化成人甲状旁腺组织,进行原代和传代细胞培养,隔日观察细胞的形态变化。取传代一代细胞,一部分通过电子显微镜观察细胞形态并检测培养液中甲状旁腺激素(parathyroid hormone,PTH)含量,另一部分采用聚合酶链反应(polymerase chain reaction,PCR)方法检测甲状旁腺细胞特异性标志物的RNA:钙敏感性受体(calcium-sensing receptor,CaSR)、PTH和神经胶质细胞缺失转录因子(glial cells missing-2,GCM-2)。结果电子显微镜下观察传代至第7 d的细胞,发现细胞质丰富,含有丰富的线粒体、内质网和高尔基体,细胞质和细胞间可见分泌囊泡;细胞培养液上清中可检测出PTH;甲状旁腺细胞特异性标志物CaSR、PTH及GCM2呈阳性。结论经胶原酶消化法消化成人甲状旁腺细胞并培养,所得甲状旁腺细胞具有良好的形态和分泌功能,并保持甲状旁腺细胞的特异表型,可作为永久性甲状旁腺功能低下时同种异体细胞移植的来源。
Objective To explore the feasibility of adult parathyroid cell culture method. Methods Adult parathyroid tissue was digested with collagenase and primary and subcultured cells were cultured. The morphological changes of the cells were observed every other day. The passage cells were passaged, the morphology of parathyroid hormone (PTH) was detected by electron microscopy and the content of parathyroid hormone (PTH) in the culture fluid. The other part was detected by polymerase chain reaction (PCR) method to detect the parathyroid cell specificity Marker RNA: calcium-sensing receptor (CaSR), PTH and glial cells missing-2 (GCM-2). Results The cells which were passaged to the 7th day under the electron microscope were found to be rich in cytoplasm, abundant in mitochondria, endoplasmic reticulum and Golgi bodies, and secreted vesicles were seen in the cytoplasm and cells; PTH was detected in the supernatant of the cell culture; Parathyroid specific markers CaSR, PTH and GCM2 were positive. Conclusion Adult parathyroid cells are digested by collagenase digestion method and cultured. The obtained parathyroid cells have good morphological and secretory functions, and maintain the specific phenotype of parathyroid cells, which can be used as the permanent allogeneic cells of hypoparathyroidism Source of transplantation.