Neural Differentiation of Human Embryonic Stem Cells at the Ultrastructural Level

Mumaw, Jennifer; Machacek, Dave W.; Shields, Joel; Dodla, Mahesh C.; Dhara, Sujoy K.; Stice, S.L.

doi:10.1017/s1431927609991279

Cited by 13 publications

(8 citation statements)

References 26 publications

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“…NP cells differentiate toward postmitotic neurons upon FGF2 withdrawal, and this event is accompanied by an initial cell death in these cultures [29]. To test the predicted role of LIF in preventing cell death during differentiation, NP cell populations differentiated in the presence of 10 ng/ml (LIF þ ) or absence (LIF À ) of LIF for 2 weeks were compared for apoptosis using TUNEL assay.…”

Section: Presence Of Lif Reduces Cell Death In Np Cell-derived Neuronsmentioning

confidence: 99%

Neurotrophic Effects of Leukemia Inhibitory Factor on Neural Cells Derived from Human Embryonic Stem Cells

et al. 2012

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Various growth factor cocktails have been used to proliferate and then differentiate human neural progenitor (NP) cells derived from embryonic stem cells (ESC) for in vitro and in vivo studies. However, the cytokine leukemia inhibitory factor (LIF) has been largely overlooked. Here, we demonstrate that LIF significantly enhanced in vitro survival and promoted differentiation of human ESC-derived NP cells. In NP cells, as well as NP-derived neurons, LIF reduced caspase-mediated apoptosis and reduced both spontaneous and H 2 O 2 -induced reactive oxygen species in culture. In vitro, NP cell proliferation and the yield of differentiated neurons were significantly higher in the presence of LIF. In NP cells, LIF enhanced cMyc phosphorylation, commonly associated with self-renewal/proliferation. Also, in differentiating NP cells LIF activated the phosphoinositide 3-kinase and signal transducer and activator of transcription 3 pathways, associated with cell survival and reduced apoptosis. When differentiated in LIF 1 media, neurite outgrowth and ERK1/2 phosphorylation were potentiated together with increased expression of gp130, a component of the LIF receptor complex. NP cells, pretreated in vitro with LIF, were effective in reducing infarct volume in a model of focal ischemic stroke but LIF did not lead to significantly improved initial NP cell survival over nontreated NP cells. Our results show that LIF signaling significantly promotes human NP cell proliferation, survival, and differentiation in vitro. Activated LIF signaling should be considered in cell culture expansion systems for future human NP cell-based therapeutic transplant studies.

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Section: Presence Of Lif Reduces Cell Death In Np Cell-derived Neuronsmentioning

confidence: 99%

Neurotrophic Effects of Leukemia Inhibitory Factor on Neural Cells Derived from Human Embryonic Stem Cells

et al. 2012

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“…Usually, differentiation is induced by EB formation followed by replating cells on an adherent system. In the replated EB center, developing columnar cells express markers of NSCs like nestin and musashi-1 and become positive for PAX6 (paired box 6a, a key transcription factor in neurogenesis, also expressed in NSCs) [136,229,228]. Over the following days the columnar cells organize into neural-tube like cells and express SOX1…”

Section: Neuronal Differentiationmentioning

confidence: 99%

Developmental and Functional Nature of Human iPSC Derived Motoneurons

Stockmann

Linta

Föhr

et al. 2011

Stem Cell Rev and Rep

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“…The ultrastructural features of high nucleocytoplasmic ratio, condensed heterochromatin and immature organelles (melanosomes and mitochondria) indicate a poorly differentiated status and suggest a link with stem-like melanoma cells. [26][27][28] This association is further supported by little or no expression of the melanocytic marker MITF 29 and the unique and high expression of FN1 indicative of an epithelial-to-mesenchymal transition-linked phenotype. 7 Furthermore, gene expression analysis of microdissected FN1 high melanoma cells revealed expression of other epithelial-to-mesenchymal transition (SNAI1 and ZEB1) and previously proposed stem-like melanoma cell markers (ABCB5, HIF2A, JARID1B, and NGFR).…”

Section: Discussionmentioning

confidence: 88%

A novel hypoxia-associated subset of FN1highMITFlow melanoma cells: identification, characterization, and prognostic value

et al. 2014

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In many human cancers, the epithelial-to-mesenchymal transition has an important role in the induction of cancer stem-like cells, and hence, in the causation of intratumoral heterogeneity. This process, also referred to as mesenchymal mimicry, is, however, only poorly understood in melanoma and histological correlation is still lacking. In an immunohistochemical analysis of a large prospective series of 220 primary and metastatic melanomas for the well-known epithelial-to-mesenchymal transition marker FN1, we observed melanoma cells with high FN1 expression in metastases with ischemic necrosis, but rarely or not at all in samples lacking evidence of hypoxia. In a blinded, retrospective series of 82 melanoma metastases with 10-year follow-up, the presence of clusters of these FN1 high melanoma cells correlated significantly with shortened melanoma-specific survival, highlighting the prognostic value of their presence. We describe in detail the unique light-and electron-microscopic features of these FN1 high melanoma cells, enabling their identification in routinely hematoxylin-and-eosin-stained sections. In addition, by laser microdissection and subsequent gene expression analysis and immunohistochemistry, we highlight their distinctive, molecular phenotype that includes expression of various markers of the epithelial-to-mesenchymal transition (eg, ZEB1) and of melanoma stemlike cells (eg, NGFR), and lack of immunoreactivity for the melanocytic marker MITF. This phenotype could be reproduced in vitro by culturing melanoma cells under hypoxic conditions. Functionally, the hypoxic microenvironment was shown to induce a more migratory and invasive cell type. In conclusion, we identified a novel clinically relevant FN1 high MITF low cell type in melanoma associated with ischemic necrosis, and propose that these cells reside at the crossroad of the epithelial-to-mesenchymal transition and stem-like cell induction, plausibly triggered by the hypoxic environment. Modern Pathology (2014) 27, 1088-1100; doi:10.1038/modpathol.2013.228; published online 3 January 2014Keywords: fibronectin (FN1); epithelial-to-mesenchymal transition; melanoma; microphthalmia-associated transcription factor (MITF); stem-like cancer cell Cutaneous melanoma is notorious for its heterogeneity, both at morphologic and phenotypic levels. 1,2 In contrast to the homogeneous appearance of in vitro melanoma cell lines, the remarkable heterogeneity of in vivo cutaneous melanoma suggests an important role of the tumor microenvironment in determining the morphology and phenotype and, consequently, the function of

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Neural Differentiation of Human Embryonic Stem Cells at the Ultrastructural Level

Cited by 13 publications

References 26 publications

Neurotrophic Effects of Leukemia Inhibitory Factor on Neural Cells Derived from Human Embryonic Stem Cells

Neurotrophic Effects of Leukemia Inhibitory Factor on Neural Cells Derived from Human Embryonic Stem Cells

Developmental and Functional Nature of Human iPSC Derived Motoneurons

A novel hypoxia-associated subset of FN1highMITFlow melanoma cells: identification, characterization, and prognostic value

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