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. 2020 Jul 15;11:446. doi: 10.3389/fendo.2020.00446

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Copyright © 2020 Ma, Shi, Morshed, Latif and Davies.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of differentiated thyroid cells. (A) qPCR analysis of thyroid specific genes: TG, TPO, TSHR, and NIS. Fold change is represented as the mean ± SEM of three independent experiments on differentiated LF cells at 21 days. (B–F) Immunostaining of thyroid genes in undifferentiated and differentiated LF cells at 21 days culture: (B,D) Undifferentiated cells as a control. (C) Staining of TG (Red) and PAX8 (Green) in differentiated cells. TG expressed in cytoplasm and PAX8 expressed in nucleus. (E) Staining of TG (Red) and NIS (Green) in a differentiated thyroid follicle: NIS was expressed in the membrane and TG was expressed in the cytoplasm and follicular lumen. (F) Thyroid neo-follicles derived from differentiated cells expressing TSHR (Green) in the membrane. Inset shows staining of rat FRTL-5 thyroid cells. Scale bar = 20 μm. (G) Measurement of T4 from the differentiated LF cells: T4 was detected in the iodine supplemented medium of the differentiated LF cells at 23 days culture as described in Methods and was absent in the medium of the undifferentiated cells but less than in FRTL-5 cells with 7H medium.