成人甲状旁腺细胞的培养和鉴定  

Culture and identification of adult human parathyroid cells

作　　者：赵越[1,2] 王晓辉[1] 刘爽[1] 孙海晨[1] 罗斌[1,3] 

机构地区：[1]首都医科大学宣武医院普通外科,北京100053 [2]山西省第二人民医院普通外科,太原030012 [3]北京清华长庚医院普通外科,清华大学医学中心,北京102218

出　　处：《中国普外基础与临床杂志》2017年第1期32-36,共5页Chinese Journal of Bases and Clinics In General Surgery

基　　金：北京市卫生系统高层次卫生技术人才项目资助(项目编号:2013-2-032);首都医科大学基础-临床科研合作基金(项目编号:14JL23)

摘　　要：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 investigate the reliability of culture method of adult human parathyroid cells. Methods Adult human parathyroid tissue was digested by collagenase, then the original generation of cells were cultured and passaged, and their morphological changes were observed and recorded every other day. Part of the passaged cells were observed through electron microscope and its supernatant parathyroid hormone （PTH） was assayed. Meanwhile, the other part of cells were tested the parathyroid markers, including PTH, calcium-sensing receptor （CaSR） and glial cells missing-2 （GCM-2） by PCR. Results Abundant cytoplasm, mitochondria, endoplasmic reticulum and Golgi apparatus from the seventh day＇s passaged cells were observed by the electron microscopy, as well as, some secretory granules existing in both cytoplasm and intercellular lacuna. Also, the PTH from supernate was detected, and parathyroid specific markers, such as CaSR, PTH, and GCM2 were positive. Conclusions These trials demonstrated the adult human parathyroid cells could be harvested by collagenase digestion and the cultured. Furthermore, the cells remained good shape and kept functioning, making it a potential source for allogeneic cell transplantation to the treatment of permanent hypoparathyroidism.

关 键 词：甲状旁腺细胞 细胞培养 聚合酶链反应 

分 类 号：R582[医药卫生—内分泌]