Generation of Thyroid Follicular Cells from Pluripotent Stem Cells: Potential for Regenerative Medicine

Sewell, Will; Lin, Reigh-Yi

doi:10.3389/fendo.2014.00096

Cited by 10 publications

(5 citation statements)

References 51 publications

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“…The subsequent manipulation of mES cells has taught us that the first step to programming such cells into a TFC pathway successfully is to lead them to form definitive endoderm. There was subsequently a series of experiments with mouse ES cells trying to obtain fully differentiated ES thyroid follicles (12). Initially, it was found that Activin A, TSH, and IGF1 were capable of inducing some TG expression but only at low levels (2,13).…”

Section: Discussionmentioning

confidence: 99%

Human Embryonic Stem Cells Form Functional Thyroid Follicles

Latif

Davies

2015

Thyroid

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Objective: The molecular events that lead to human thyroid cell speciation remain incompletely characterized. It has been shown that overexpression of the regulatory transcription factors Pax8 and Nkx2-1 (ttf-1) directs murine embryonic stem (mES) cells to differentiate into thyroid follicular cells by initiating a transcriptional regulatory network. Such cells subsequently organized into three-dimensional follicular structures in the presence of extracellular matrix. In the current study, human embryonic stem (hES) cells were studied with the aim of recapitulating this scenario and producing functional human thyroid cell lines. Methods: Reporter gene tagged pEZ-lentiviral vectors were used to express human PAX8-eGFP and NKX2-1-mCherry in the H9 hES cell line followed by differentiation into thyroid cells directed by Activin A and thyrotropin (TSH). Results: Both transcription factors were expressed efficiently in hES cells expressing either PAX8, NKX2-1, or in combination in the hES cells, which had low endogenous expression of these transcription factors. Further differentiation of the double transfected cells showed the expression of thyroid-specific genes, including thyroglobulin (TG), thyroid peroxidase (TPO), the sodium/iodide symporter (NIS), and the TSH receptor (TSHR) as assessed by reverse transcription polymerase chain reaction and immunostaining. Most notably, the Activin/ TSH-induced differentiation approach resulted in thyroid follicle formation and abundant TG protein expression within the follicular lumens. On stimulation with TSH, these hES-derived follicles were also capable of dosedependent cAMP generation and radioiodine uptake, indicating functional thyroid epithelial cells. Conclusion: The induced expression of PAX8 and NKX2-1 in hES cells was followed by differentiation into thyroid epithelial cells and their commitment to form functional three-dimensional neo-follicular structures. The data provide proof of principal that hES cells can be committed to thyroid cell speciation under appropriate conditions.

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Section: Discussionmentioning

confidence: 99%

Human Embryonic Stem Cells Form Functional Thyroid Follicles

Latif

Davies

2015

Thyroid

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“…Thyroid deficiency is common and about 12% of the population in US may suffer the thyroid related conditions during lifetime (Sewell et al, 2014;Agrawal et al, 2015). There have been lots of studies about stem cell technologies which can be the key tools for tissue engineering and regenerative medicine to treat this condition in the recent years.…”

Section: Discussionmentioning

confidence: 99%

“…Thyroid gland produces two essential hormones which impact almost every organ and cell of the body and get involved in varieties of biological processes (Sewell et al, 2014). Hypothyroidism, an irreversible change (Ma et al, 2015), is the most common hormonal deficiency and can be precipitated by various diseases (Hollowell et al, 2002;McLachlan et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Hypoxia promotes thyroid differentiation of native murine induced pluripotent stem cells

Yang

Lu²,

Chen³

et al. 2016

Int. J. Dev. Biol.

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Hypothyroidism is a very common hormonal deficiency and the stem cell technology which developed in the recent years may offer a therapeutic strategy for treating this disorder. Hypoxia has been demonstrated to play an important role in embryonic formation and development and to modulate stem cell differentiation. However, the influence of oxygen tension on thyroid differentiation has not been studied. In this study, we used murine induced pluripotent stem (iPS) cells for thyroid cell differentiation under normoxic and hypoxic conditions and compared differentiation efficiency in morphology, function, gene and protein expression under both conditions. We found that hypoxia promoted adhesion and outgrowth of embryoid bodies (EBs) derived from murine iPS cells. Expression of endodermal markers (Foxa2 and Gata4) and thyroid transcription factors (Pax8 and Nkx2.1) was increased by hypoxia at both gene and protein levels during earlymid differentiation stages (p<0.05). And so were the thyroid specific markers NIS and TSHR at the end of the experiment (p<0.05). In addition, functional iodide uptake by differentiated cells was also increased after hypoxia. Thyroid differentiation from iPS cells is enhanced under hypoxia and this may involve hypoxia inducible factors (HIFs) and their downstream gene FGF2. Our data offer a foundation for understanding thyroid development and provide a potentially more efficient way to use cell therapy for treating thyroid deficiency.

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“…The reprogrammed cells can then be differentiated into any desired cell type and used in cell-based therapy, disease modeling or for drug discovery. Picture adapted from (100).…”

Section: Figure 2 Induced Pluripotent Stem Cellmentioning

confidence: 99%

Passaging Techniques and ROCK Inhibitor Exert Reversible Effects on Morphology and Pluripotency Marker Gene Expression of Human Embryonic Stem Cell Lines

Holm

Nikdin²,

Kjartansdóttir³

et al. 2013

Stem Cells and Development

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Human embryonic stem cells (hESCs) are known for their potential usage in regenerative medicine, but also for handling sensitivity. Much effort has been put into optimizing the culture methods of hESCs. It has been shown that the use of Rho-associated coiled-coil kinase inhibitor (ROCKi) decreases the cellular stress response and the apoptotic cell death in hESC cultures that have been passaged enzymatically. These observations sparked a wide use of ROCKi in hESC cultures. We and others, however, noted that cells passaged enzymatically with the use of ROCKi had a different morphology compared to cells passaged mechanically. Here we show that hESCs that were enzymatically passaged displayed alterations in the nuclear size compared to cultures that were mechanically passaged. Notably, a dramatically decreased expression of the genes encoding common pluripotency markers, such as OCT4/POU5F1 and NANOG were revealed in enzymatically passaged hESCs compared to mechanically passaged, while such differences were not significant when assessing protein levels. The differences in gene expression did not correlate strongly with commonly analyzed histone modifications (H3K4me3, H3K9me3, H3K27me3, and H4K16ac) on the promoters of these genes. Surprisingly, the effects of enzymatic passaging were at least in part reversible as the gene expression profile of enzymatically passaged hESCs that were transferred back to mechanical passaging, showed no significant difference compared to those hESCs that were continuously passaged mechanically. Our results suggest that enzymatic passaging influences parameters associated with hESC characteristics, and emphasizes the importance of using cells handled in the same manner when comparing results both within and between projects.

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Generation of Thyroid Follicular Cells from Pluripotent Stem Cells: Potential for Regenerative Medicine

Cited by 10 publications

References 51 publications

Human Embryonic Stem Cells Form Functional Thyroid Follicles

Human Embryonic Stem Cells Form Functional Thyroid Follicles

Hypoxia promotes thyroid differentiation of native murine induced pluripotent stem cells

Passaging Techniques and ROCK Inhibitor Exert Reversible Effects on Morphology and Pluripotency Marker Gene Expression of Human Embryonic Stem Cell Lines

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