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[人甲状腺结节组织中甲状腺干细胞的分离、增殖与分化]

[Isolation, proliferation and differentiation of thyroid stem cell from human thyroid nodule tissue].

作者信息

Lan Ling, Cui Dai, Shi Bing-yin, Zhang Guo-ying, Deng Wei, Wang Hong, Chen Hai-ling, Deng Li-li

机构信息

Department of Endocrinology, Beijing Jishuitan Hospital, Fourth Medical College of Peking University, Beijing 100035, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2012 Mar 27;92(12):806-10.

Abstract

OBJECTIVE

To explore the presence of a characteristic stem cell population (side population, SP) in human thyroid gland and perform sphere culture method for the isolation and proliferation of thyroid stem cell.

METHODS

Flow cytometry and cell sorting were performed to identify and isolate the ABCG2-positive SP cells from primary thyroid cells. The comparison of gene profiles and morphology between SP and main population (MP) were performed. Primary thyroid cells were also cultured in neurosphere-like growth condition for sphere formation. Gene profile of developed spheres was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and thyroid lineage commitment was then induced in a differentiating condition. In stem cell-derived thyrocytes, embedded in collagen to form follicles, TSH-dependent (125)iodide uptake was measured.

RESULTS

The SP was identified as a population enriched in stem cells with typical morphology, and characteristics of self-renewal and differentiation. Nonadherent clonal spheres developed in thyroid cell cultures, displaying an expression pattern resembling that of SP cells and in response to TSH and serum these sphere cells differentiated into thyrocytes expressing PAX8, thyroglobulin, sodium iodide symporter, thyroid-stimulating hormone receptor and thyroperoxidase mRNA. And there was TSH-dependent (125)iodide uptake.

CONCLUSION

It is shown first time that human thyroid contains an undescribed population of cells with SP phenotype and clonal expansion capacity. Moreover sphere culture method is developed for the isolation and proliferation of thyroid stem cell.

摘要

目的

探索人甲状腺中特征性干细胞群(侧群细胞,SP)的存在,并采用球囊培养法分离和增殖甲状腺干细胞。

方法

运用流式细胞术和细胞分选技术从原代甲状腺细胞中鉴定并分离出ABCG2阳性的SP细胞。对SP细胞和主要细胞群(MP)的基因谱和形态进行比较。原代甲状腺细胞也在类神经球生长条件下进行培养以形成球囊。通过逆转录聚合酶链反应(RT-PCR)确定所形成球囊的基因谱,然后在分化条件下诱导甲状腺谱系定向分化。在包埋于胶原中形成滤泡的干细胞来源的甲状腺细胞中,检测促甲状腺激素(TSH)依赖性碘摄取。

结果

SP细胞被鉴定为富含具有典型形态、自我更新和分化特征的干细胞群。甲状腺细胞培养中形成了非贴壁克隆球囊,其表达模式类似于SP细胞,并且在TSH和血清作用下,这些球囊细胞分化为表达PAX8、甲状腺球蛋白、钠碘同向转运体、促甲状腺激素受体和甲状腺过氧化物酶mRNA的甲状腺细胞。并且存在TSH依赖性碘摄取。

结论

首次表明人甲状腺中含有具有SP表型和克隆扩增能力的未被描述的细胞群。此外,开发了球囊培养法用于甲状腺干细胞的分离和增殖。

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