Cell cycle synchronization of embryonic stem cells: Effect of serum deprivation on the differentiation of embryonic bodies in vitro

Zhang, Enming; Li, Xiaolong; Zhang, Shufang; Chen, Liangqiang; Zheng, Xiaoxiang

doi:10.1016/j.bbrc.2005.05.200

Cited by 50 publications

(42 citation statements)

References 17 publications

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“…Mouse ES-pS3/RR cells were starved of serum and LIF for 24 h followed by stimulation with 10 3 U/mL of LIF for the indicated times. Similar to the study by Zhang et al [27], most of these cells were still pluripotent and could be used as a model for STAT3 signaling. During the LIF withdrawal period, the cells retained ES cell-like morphology.…”

Section: Stat3 Activation Is Involved In the Lif Pathwaymentioning

confidence: 70%

Imaging of STAT3 Signaling Pathway During Mouse Embryonic Stem Cell Differentiation

Xie

Chan²,

Cao³

et al. 2009

Stem Cells and Development

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Signal transducers and activators of transcription 3 (STAT3) is a pleiotropic transcription factor involved in a variety of physiological processes. STAT3 acts as a key transcriptional determinant of mouse embryonic stem (ES) cell self-renewal and plays a pivotal function in early mammalian embryogenesis because the development of many organs requires STAT3 activation. However, little is known about the role of STAT3 function during ES cell differentiation. To answer this question, we built a lentiviral construct with 7-repeat STAT3-binding sequence (enhancer) and minimal TA (promoter) driving renilla luciferase and monomeric red fl uorescence protein (Rluc-mRFP), followed by a constitutive cytomegalovirus promoter driving green fl uorescent protein as a selection marker. The specifi city of our custom-designed 7-repeat STAT3 reporter construct was fi rst confi rmed by cotransfection with constitutively active version of STAT3 (STAT3C) into human embryonic kidney 293T cells. Next, a mouse ES cell line stably transduced with STAT3 reporter construct was isolated. This ES cell line showed a tight response in reporter gene expression with leukemia inhibitory factor (LIF) induction and was chosen as a developmental model for the STAT3 functional study. Using serial noninvasive bioluminescence imaging, we showed that the onset of embryoid body (EB) formation involved inhibition of STAT3 activity. However, during differentiation, STAT3 activity steadily increased from day 5 to 14 and was reduced by day 21. STAT3 activity was also confi rmed separately by Western blots. Finally, phosphorylation of STAT3 was also found to correspond with cardiomyocyte differentiation. In summary, this is the fi rst study to monitor real-time STAT3 activity during ES cell differentiation. This genetically modifi ed line can be used to study the biological role of STAT3 during ES cell differentiation into different derivatives.

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Section: Stat3 Activation Is Involved In the Lif Pathwaymentioning

confidence: 70%

Imaging of STAT3 Signaling Pathway During Mouse Embryonic Stem Cell Differentiation

Xie

Chan²,

Cao³

et al. 2009

Stem Cells and Development

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“…The [ 3 H]thymidine incorporation experiments were carried out using the methodology described by Brett et al (2). Zhang et al (41) reported that most ES cells could be arrested in the G 0/G1 phase through serum deprivation. Furthermore, the synchronized ES cells could successfully reenter a normal cell cycle after resupplying the serum.…”

Section: Materials Mouse Es Cells Were Obtained From American Type Cmentioning

confidence: 99%

Arachidonic acid potentiates hypoxia-induced VEGF expression in mouse embryonic stem cells: involvement of Notch, Wnt, and HIF-1α

Lee¹,

Kim²,

Han³

2009

American Journal of Physiology-Cell Physiology

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“…Embryonic bodies (EBs) were formed as described previously (Zhang et al, 2005). To differentiate EBs into neuronal cells, retinoic acid (5 M) was treated for 4 days (Okada et al, 2004).…”

Section: Embryonic Body Formation and Neuronal Differentiationmentioning

confidence: 99%

Dynamic changes of gangliosides expression during the differentiation of embryonic and mesenchymal stem cells into neural cells

Kwak

Yu²,

Kim³

et al. 2006

Exp Mol Med

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Stem cells are used for the investigation of developmental processes at both cellular and organism levels and offer tremendous potentials for clinical applications as an unlimited source for transplantation. Gangliosides, sialic acid-conjugated glycosphingolipids, play important regulatory roles in cell proliferation and differentiation. However, their expression patterns in stem cells and during neuronal differentiation are not known. Here, we investigated expression of gangliosides during the growth of mouse embryonic stem cells (mESCs), mesenchymal stem cells (MSCs) and differentiated neuronal cells by using high-performance thin-layer chromatography (HPTLC). Monosialoganglioside 1 (GM1) was expressed in mESCs and MSCs, while GM3 and GD3 were expressed in embryonic bodies. In the 9-day old differentiated neuronal cells from mESCs cells and MSCs, GM1 and GT1b were expressed. Results from immunostaining were consistent with those observed by HPTLC assay. These suggest that gangliosides are specifically expressed according to differentiation of mESCs and MSCs into neuronal cells and expressional difference of gangliosides may be a useful marker to identify differentiation of mESCs and MSCs into neuronal cells.

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Cell cycle synchronization of embryonic stem cells: Effect of serum deprivation on the differentiation of embryonic bodies in vitro

Cited by 50 publications

References 17 publications

Imaging of STAT3 Signaling Pathway During Mouse Embryonic Stem Cell Differentiation

Imaging of STAT3 Signaling Pathway During Mouse Embryonic Stem Cell Differentiation

Arachidonic acid potentiates hypoxia-induced VEGF expression in mouse embryonic stem cells: involvement of Notch, Wnt, and HIF-1α

Dynamic changes of gangliosides expression during the differentiation of embryonic and mesenchymal stem cells into neural cells

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