Conversion of rat embryonic stem cells into neural precursors in chemical-defined medium

Peng, Xinrong; Gao, Huile; Wang, Ying; Yang, Baotian; Liu, Tao; Sun, Yawang; Jin, Haixia; Jiang, Lihua; Li, Linfang; Wu, Mengchao; Qian, Qijun

doi:10.1016/j.bbrc.2013.01.023

Cited by 6 publications

(8 citation statements)

References 21 publications

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“…Furthermore, a tumor cell line was established by culturing cells in a chemically defined N2B27 medium containing CHIR99021. The cell lines revealed some indications of malignancy, such as Oct4 expression and a long-term expansion ability (20).…”

Section: Discussionmentioning

confidence: 99%

GSK‑3 inhibitor CHIR99021 enriches glioma stem‑like cells

Yang

Wang²,

Bozinov

et al. 2020

Oncol Rep

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Glioblastoma (GBM) is the most prevalent and lethal primary intrinsic brain cancer. The disease is essentially incurable, with glioblastomas characterized by resistance to both chemotherapy and radiotherapy, as well as by rapid tumor progression, all of which are mainly ascribed to glioma stem-like cells (GSLCs). In the present study, an improved model that is more similar to clinical GBM was constructed. Twenty clinical glioma samples were collected to obtain primary low-grade tumor cells. The cells were either maintained in serum-free medium as primary glioma-based cells (PGBCs) or cultured in the same medium with CHIR99021 as GSLCs. Then, the molecular and ultrastructural differences between the two cell groups were determined. Furthermore, the proliferation and migration of the GSLCs were examined and the potential mechanisms were investigated. Finally, temozolomide resistance in vitro and in the mouse model was assessed to study the properties of the induced GSLCs. The primary low-grade tumor cells extracted from surgical samples were enriched with GSLC properties, with high expression levels of CD133 and Nestin in 100 nM CHIR99021. The GSLCs exhibited high proliferation and migration. Furthermore, the expression of the PI3K/AKT signaling pathway and that of related genes and proteins were significantly enhanced by CHIR99021. The animal study also revealed high levels of STAT3, mTOR, NF-κB, and VEGF in the GSLC-transplanted mice. CHIR99021 could stably enhance GSLC properties in patient-derived glioma samples. It may provide a useful model for further study, helping to understand the pathogenesis of therapeutic resistance and to screen drug candidates.

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Section: Discussionmentioning

confidence: 99%

GSK‑3 inhibitor CHIR99021 enriches glioma stem‑like cells

Yang

Wang²,

Bozinov

et al. 2020

Oncol Rep

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“…Despite of the growing list of differentiation protocols for mESCs, differentiation of rESCs is extremely difficult to achieve in vitro . To date, only two strategies have been described in which rESCs were triggered to differentiate into either cardiomyocytes or neuronal precursors and in these differentiation protocols MEK and GSK3β inhibitors, that are commonly used for ESC culture, are always present in low concentrations in the differentiation media 74,75 . XCI is closely linked to loss of pluripotency, and the presence of an inactive X chromosome provides a powerful readout for cell differentiation.…”

Section: Resultsmentioning

confidence: 99%

A novel approach to differentiate rat embryonic stem cells in vitro reveals a role for RNF12 in activation of X chromosome inactivation

Magaraki

Loda

Gontan

et al. 2019

Sci Rep

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X chromosome inactivation (XCI) is a mammalian specific, developmentally regulated process relying on several mechanisms including antisense transcription, non-coding RNA-mediated silencing, and recruitment of chromatin remodeling complexes. In vitro modeling of XCI, through differentiation of embryonic stem cells (ESCs), provides a powerful tool to study the dynamics of XCI, overcoming the need for embryos, and facilitating genetic modification of key regulatory players. However, to date, robust initiation of XCI in vitro has been mostly limited to mouse pluripotent stem cells. Here, we adapted existing protocols to establish a novel monolayer differentiation protocol for rat ESCs to study XCI. We show that differentiating rat ESCs properly downregulate pluripotency factor genes, and present female specific Xist RNA accumulation and silencing of X-linked genes. We also demonstrate that RNF12 seems to be an important player in regulation of initiation of XCI in rat, acting as an Xist activator. Our work provides the basis to investigate the mechanisms directing the XCI process in a model organism different from the mouse.

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“…However, in rodent-based models, this was inconsistent. Indeed, two studies reported that in rat-derived ESCs and in mouse embryonic fibroblast, CHIR promoted neural differentiation similar to the human model [37,38], but two other studies showed that in mouse ESCs CHIR inhibits differentiation, enhancing pluripotency-maintenance mechanisms [39,40]. This discrepancy between human and murine in-vitro platforms, highlights the importance of the use of human-based models in neurodevelopmental research.…”

Section: Discussionmentioning

confidence: 99%

Neural differentiation is increased by GSK-3β inhibition and decreased by tankyrase inhibition in human neural precursor cells

Telias

Ben‐Yosef

2019

Preprint

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Glycogen synthase kinase-3β (GSK-3β) and tankyrase-1/2 (TANK) are two enzymes known to play multiple roles in cell biology, including regulation of proliferation, differentiation and metabolism. Both of them act on the canonical Wnt/β-Catenin pathway, but are also involved in many other independent intracellular mechanisms.More importantly, GSK-3β and TANK have been shown to play crucial roles in different diseases, including cancer and neurological disorders. The GSK-3β-inhibitor 'CHIR' and the TANK-inhibitor 'XAV' are two pyrimidine molecules, holding high potential as possible therapeutic drugs. However, their effect on neural tissue is poorly understood. In this study, we tested the effects of CHIR and XAV on human neural precursor cells (hNPCs) derived from human embryonic stem cells. We found that CHIR-mediated inhibition of GSK-3β promotes neural differentiation. In contrast, XAV-mediated inhibition of TANK leads to de-differentiation. These results highlight the relative importance of these two enzymes in determining the neurodevelopmental status of hNPCs. Furthermore, they shed light on the roles of Wnt signaling during early human neurogenesis.

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Conversion of rat embryonic stem cells into neural precursors in chemical-defined medium

Cited by 6 publications

References 21 publications

GSK‑3 inhibitor CHIR99021 enriches glioma stem‑like cells

GSK‑3 inhibitor CHIR99021 enriches glioma stem‑like cells

A novel approach to differentiate rat embryonic stem cells in vitro reveals a role for RNF12 in activation of X chromosome inactivation

Neural differentiation is increased by GSK-3β inhibition and decreased by tankyrase inhibition in human neural precursor cells

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