人甲状腺原代细胞的长期培养  

Long-term primary culture of human thyroid cells

作　　者：刘东方[1] 邓华聪[1] 许改平[2] 雄波 侯平 

机构地区：[1]重庆医科大学附属一院内分泌科,400016 [2]重庆医科大学附二院内分泌科 [3]重庆壁山县人民医院外科

出　　处：《中华内分泌代谢杂志》2005年第2期181-183,共3页Chinese Journal of Endocrinology and Metabolism

基　　金：重庆市卫生局(2001 1 15)

摘　　要：目的　探索人甲状腺细胞长期原代培养的方法及细胞在不同时期的功能变化。方法　常规分离甲状腺细胞,并用含有10%胎牛血清(FBS)、谷氨酰胺、牛胰岛素、10mU/LTSH和氢化可的松的MEM培养,细胞铺成单层后换用4% FBC的培养基培养,分别在不同时间观察甲状腺细胞吸碘率、细胞生长情况,免疫组化观察甲状腺特异抗原甲状腺球蛋白(Tg)表达水平,RT PCR测定钠碘转运子(NIS)基因表达水平。结果　甲状腺细胞在培养40d左右仍生长良好,培养14d细胞具有80%的吸碘率,培养7dTg表达可达95%,培养14dTg表达仍达60%,NISmRNA表达在培养7d达98%,而培养14d表达率只有40%。结论　应用该研究所建立的培养条件可以使甲状腺细胞生存40d以上,该实验条件下培养3, 7, 14,28d的原代甲状腺滤泡上皮细胞以培养7d的细胞NISmRNA,Tg表达、摄碘功能最接近基线水平。Objective To investigate the primary culture method of human thyroid follicular cells for a long term and the function of these cells cultured for different days. Methods Thyroid cells were isolated following routine procedures and cultured in the MEM supplemented with 10% fetal bovine serum (FBS), glutamine, hydrocortisone, insulin, TSH and T_ 3. When thyroid cells grew into a monolayer, the culture medium was replaced by the medium with 4% FBS. Iodide uptake rate, cell morphology, specific antigen (thyroglobulin, Tg) expression and gene expression of sodium iodide symporter (NIS) of thyroid cells cultured for different days were observed. Tg antigen expression and NIS mRNA expression were observed by immunohistochemistry and RT-PCR, respectively. Results The thyroid cells grew satisfactorily up to 40 days of incubation. There was a 80% iodide uptake rate by the cultured cells within 14 days. Tg expression of thyroid cells was 95% on 7th day and 60% on 14th day. NIS mRNA relative expression of thyroid cells was 98% on the 7th day,only 40% on the 14th day. Conclusion The thyroid cells cultured with present method are growing up to over 40 days, and the cells cultured on the 7th day preserves the closest NIS gene、TG antigen expression and iodide uptake function to baseline.

关 键 词：甲状腺原代细胞 细胞培养 甲状腺疾病 谷氨酰胺 牛胰岛素 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]