Laminin switches terminal differentiation fate of human trophoblast stem cells under chemically defined culture conditions

Karakis, Victoria; Jabeen, Mahe; Britt, John; Cordiner, Abigail; Mischler, Adam; Li, Feng; Miguel, Adriana San; Rao, Balaji M.

doi:10.1016/j.jbc.2023.104650

Cited by 3 publications

(12 citation statements)

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“…We further assessed the differentiation of T1 and T2 hTSCs derived from term CTBs to the STB and EVT lineages using chemically defined differentiation conditions previously described by Karakis et al 15 , or using protocols described by Okae et al 5 We have previously described chemically defined conditions for STB differentiation of hTSCs in the absence of forskolin 15 . Under these conditions, a membrane stain showed that T1 and T2 hTSCs formed multinucleate cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…We have previously described chemically defined conditions for STB differentiation of hTSCs in the absence of forskolin 15 . Under these conditions, a membrane stain showed that T1 and T2 hTSCs formed multinucleate cells ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Image analysis was conducted using an image processing algorithm created in MATLAB R2021a, as previously described 15 . Briefly, TIFF images were generated after background correction in Zeiss Zen Lite software, using settings for corresponding isotype images for all samples.…”

Section: Confocal Microscopy Image Analysismentioning

confidence: 99%

“…The code used for image analysis can be found at https://doi.org/10.5281/zenodo.7700524. Statistical analysis was conducted using the non-parametric Mann-Whitney U test, in Microsoft Excel as described 15 and Ca 2+ ) using a Keyence BZ-X810 system. The fusion index for each condition was calculated using two biological replicates per cell line, except for T2 hTSCs in TUA medium differentiated using the protocol by Karakis et al, where a single replicate was used.…”

Section: Confocal Microscopy Image Analysismentioning

confidence: 99%

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Derivation of human trophoblast stem cells from placentas at birth

Karakis,

Britt,

Jabeen

et al. 2024

Preprint

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Human trophoblast stem cells (hTSCs) have emerged as a powerful tool for modeling the placental cytotrophoblast (CTB) in vitro. hTSCs were originally derived from CTBs of the first trimester placenta or blastocyst-stage embryos in trophoblast stem cell medium (TSCM) that contains epidermal growth factor (EGF), the glycogen synthase kinase-beta (GSK3β) inhibitor CHIR99021, the transforming growth factor-beta (TGFβ) inhibitors A83-01 and SB431542, valproic acid (VPA), and the Rho-associated protein kinase (ROCK) inhibitor Y-27632. Here we show that hTSCs can be derived from CTBs isolated from the term placenta, using TSCM supplemented with a low concentration of mitochondrial pyruvate uptake inhibitor UK5099 and lipid-rich albumin (TUA medium). Notably, hTSCs could not be derived from term CTBs using TSCM alone, or in the absence of either UK5099 or lipid-rich albumin. Strikingly, hTSCs cultured in TUA medium for a few passages could be transitioned into TSCM and cultured thereafter in TSCM. hTSCs from term CTBs cultured in TUA medium as well as those transitioned into and cultured in TSCM thereafter could be differentiated to the extravillous trophoblast and syncytiotrophoblast lineages and exhibited high transcriptome similarity with hTSCs derived from first trimester CTBs. We anticipate that these results will enable facile derivation of hTSCs from normal and pathological placentas at birth with diverse genetic backgrounds and facilitate in vitro mechanistic studies in trophoblast biology.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Confocal Microscopy Image Analysismentioning

confidence: 99%

Section: Confocal Microscopy Image Analysismentioning

confidence: 99%

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