Normal Human Thyrocytes in Culture.
Normal Human Thyrocytes in Culture.
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DOI：
10.1007/978-1-4939-8600-2_1
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Gershengorn MC
中科院分区：
文献类型：
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作者：
Morgan SJ;Neumann S;Gershengorn MC
为了研究正常人甲状腺细胞的功能，我们开发了一种在原代培养中获得这些细胞的方案。甲状腺细胞获自手术切除甲状腺肿瘤时获得的正常组织。在无菌条件下，将样本切碎成小块，通过用IV型胶原酶消化产生单分散的细胞，并将细胞接种在含有10%胎牛血清（FBS）的Dulbecco改良Eagle培养基（DMEM）中的组织培养级培养皿中。在37 °C下在加湿的5%C02培养箱中孵育24小时后，除去含有非贴壁细胞的上清液，并将贴壁细胞在含有10%FBS、100 IU/mL青霉素和10 μg/mL链霉素的DMEM中增殖。细胞以72-94小时的倍增时间增殖，并保留9-12个倍增时间的功能特征。我们已经成功地使用它们在研究中阐明促甲状腺激素（TSH）和胰岛素样生长因子1的信号转导。
In order to study functions of normal human thyrocytes, we developed a protocol to obtain these cells in primary culture. Thyrocytes are obtained from normal tissue obtained at surgery for removal of thyroid neoplasms. Under sterile conditions, specimens are minced into small pieces, mono-dispersed cells are generated by digestion with collagenase type IV and the cells plated in tissue culture grade dishes in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS). After 24 h of incubation at 37 °C in a humidified 5% CO2 incubator, the supernatant containing non-adherent cells is removed and the adherent cells are propagated in DMEM with 10% FBS, 100 IU/mL penicillin, and 10 μg/mL streptomycin. Cells proliferate with a doubling time of 72–94 h and retain functional characteristics for 9–12 doublings. We have used them successfully in studies to elucidate the signaling by thyrotropin (TSH) and insulin-like growth factor 1.