Differentiation of rhesus embryonic stem cells to neural progenitors and neurons

Calhoun, John; Lambert, Nevin A.; Mitalipova, Maisam; Noggle, Scott; Lyons, Ian; Condie, Brian G.; Stice, Steven L.

doi:10.1016/s0006-291x(03)00937-9

Cited by 32 publications

(27 citation statements)

References 41 publications

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“…The latter have been proposed to act as fate determinants in embryonic neural stem cells [29]. Neural rosettes are conserved among species [23,26,30,31] and positive for early neural markers, such as Sox1, Sox2, Nestin, and Pax6 [23,26,31]. Recently, it has been shown in human ESCs that neural rosettes represent an early neural stem cell stage [26].…”

Section: Introductionmentioning

confidence: 99%

The Apical Polarity Determinant Crumbs 2 Is a Novel Regulator of ESC-Derived Neural Progenitors

Boroviak

Rashbass

2011

Stem Cells

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ESCs undergoing neural differentiation in vitro display an intrinsic heterogeneity with a large subset of the cells forming polarized neural rosettes that maintain the neural progenitor microenvironment. This heterogeneity is not only necessary for normal development but also causes substantial technical challenges for practical applications. Here, we report a novel regulator of early neural progenitors, the apical polarity protein Crb2 (Crumbs homologue 2). Employing monolayer differentiation of mouse ESCs to model neurogenesis in vitro, we find that Crb2 is upregulated with Sox1 and Musashi at the onset of neuroepithelial specification and localizes to the apical side of neural rosettes. Stable Crb2-knockdown (KD) lines die at the onset of neural specification and fail to stabilize several apical polarity proteins. However, these cells are able to proliferate under selfrenewing conditions and can be differentiated into mesodermal and endodermal lineages. Conversely, Crb2 overexpression during neural differentiation results in elevated levels of other apical polarity proteins and increases proliferation. Additionally, sustained overexpression of Crb2 reduces terminal differentiation into TuJ1-positive neurons. Furthermore, we demonstrate that Crb2 overexpression under selfrenewing conditions increases glycogen synthase kinase (GSK)-3b inhibition, correlating with an increase in clonogenicity. To confirm the importance of GSK-3b inhibition downstream of Crb2, we show that Crb2-KD cells can be forced into neural lineages by blocking GSK-3b function and supplementing Epidermal Growth Factor (EGF) and basic Fibroblast Growth Factor (bFGF). Thus, this is the first demonstration that a member of the Crumbs family is essential for survival and differentiation of ESC-derived neural progenitors.

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Section: Introductionmentioning

confidence: 99%

The Apical Polarity Determinant Crumbs 2 Is a Novel Regulator of ESC-Derived Neural Progenitors

Boroviak

Rashbass

2011

Stem Cells

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“…Recently, various types of cells differentiated from hESCs or nonhuman ESCs have been reported [18,[26][27][28][29][30][31][32][33][34][35][36]. For example, hematopoietic cells, dopaminergic neurons, and insulin-producing cells have been generated from hESCs or primate ESCs.…”

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confidence: 99%

Establishment of Novel Embryonic Stem Cell Lines Derived from the Common Marmoset ( Callithrix jacchus )

et al. 2005

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The successful establishment of human embryonic stem cell (hESC) lines has inaugurated a new era in regenerative medicine by facilitating the transplantation of differentiated ESCs to specific organs. However, problems with the safety and efficacy of hESC therapy in vivo remain to be resolved. Preclinical studies using animal model systems, including nonhuman primates, are essential to evaluate the safety and efficacy of hESC therapies. Previously, we demonstrated that common marmosets are suitable laboratory animal models for preclinical studies of hematopoietic stem cell therapies. As this animal model is also applicable to preclinical trials of ESC therapies, we have established novel common marmoset ESC (CMESC) lines. To obtain marmoset embryos, we developed a new embryo collection system, in which blastocysts can be obtained every 3 weeks from each marmoset pair. The inner cell mass was isolated by immunosurgery and plated on a mouse embryonic feeder layer. Some of the CMESC lines were cultured continuously for more than 1 year. These CMESC lines showed alkaline phosphatase activity and expressed stage-specific embryonic antigen (SSEA)-3, SSEA-4, TRA-1-60, and TRA-1-81. On the other hand, SSEA-1 was not detected. Furthermore, our novel CMESCs are pluripotent, as evidenced by in vivo teratoma formation in immunodeficient mice and in vitro differentiation experiments. Our established CMESC lines and the common marmoset provide an excellent experimental model system for understanding differentiation mechanisms, as well as the development of regenerative therapies using hESCs. Stem Cells 2005;23:1304-1313 This material is protected by U.S.

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“…Following the retinoic acid in vitro exposure of P19 (EC cell line) and embryo derived pluripotential stem cell line, they differentiated into neural cells [10,11,34,35] . Different studies have utilized other signal molecules such as βNGF, FGF, Wnt, TGF-β and N2 with different protocols in order to promote neural differentiation [14,36,37] .…”

Section: Discussionmentioning

confidence: 99%

The Differentiation of Neuronal Cells from Mouse Embryonic Stem Cells

Umur¹,

Vatansever²,

Umur³

et al. 2014

Kafkas Univ Vet Fak Derg

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ÖzetWith new technologies emerging today, the importance of stem cells in the cell therapy of nervous system diseases is supported by recent studies. Therefore, the development of neuronal cell differentiation protocols from stem cells is of great importance. In our study, the differentiation of neuronal and neuroglial cells from mouse embryonic stem (ES) cell line and their analysis with neuronal cell markers are aimed. Mouse ES cells were differentiated to neurogenic series cells by adding N2 and bFGF to the culture medium on coated Fibronectin dishes. For the identification of differentiated cells, they were evaluated by light microscopy using immunhistochemistry techniques and by electron microscopy. Indirect immunohistochemical staining method was performed with SSEA-1 (mouse embriyonic stem cells marker), Nestin (neural precursor cells marker), βIII-Tubulin (neuronal cells marker), MAP-2 (neuronal cells marker), GFAP (astrocyte marker), and O4 (oligodendrocyte marker). After 1 week of differentiation of cells, immunoreactivities of SSEA-1 and Nestin were detected to be negative and moderate, respectively. After 2 weeks culture time, the differentiation was still continuing and especially positive immunoreactivities of β-III Tubulin and MAP-2 and weak immunoreactivities of O4 and GFAP were supported neuronal differentiation. In conclusion, our results suggest that neuronal cell derived from mouse ES cells were differentiated particularly to neuron using N2+bFGF+fibronectin culture condition. Therefore, these differentiated cells may be used as a treatment method in degenerative diseases of the nervous system. Anahtar sözcükler: Mouse embryonic stem cell, Differentiation, Neuron and neuroglia Fare Embriyonik Kök Hücrelerden Nöronal Hücrelerin Farklılaşması SummaryGünümüzde gelişen yeni teknolojiler sayesinde sinir sistemi hastalıklarının hücresel tedavisinde kök hücrelerinin önemi son yıllardaki çalışmalar ile desteklenmektedir. O nedenle kök hücrelerden nöronal hücrelerin farklılaştırılması protokollerinin oluşturulması büyük önem taşımaktadır. Çalışmamızda, fare embriyonik kök (EK) hücre hattından, nöron ve nöroglial hücrelerin farklılaştırılması ve nöronal hücre belirteçleri ile analizi amaçlanmıştır. Fare EK hücreler, fibronektin kaplı petrilerde kültür ortamına N2 ve bFGF ilavesi ile nörojenik seri hücrelerine farklılaştırıldı. Farklılaşmış hücrelerin tanımlanması için hücreler, immünohistokimya tekniği kullanılarak ışık mikroskobu ile ve elektron mikroskobu ile değerlendirildi. İndirek immünohistokimyasal boyama yöntemi SSEA-1 (fare ES hücre belirteci), Nestin (nöron öncülü hücre belirteci), βIII-Tubulin (nöron hücre belirteci), MAP-2 (nöron hücre belirteci), GFAP (astrosit belirteci) ve O4 (oligodendrosit belirteci) için uygulandı. Hücrelerin farklılaşmasının 1. haftasından sonra SSEA-1 ve Nestin immünoreaktivitesi sırasıyla negatif ve orta saptandı. Kültürün 2. haftasından sonra farklılaşmanın hala devam etmesi ve özellikle β-III Tubulin ve MAP-2 immünoreaktivitesinin güçlü pozitif ve O4 ve GFAP immü...

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Differentiation of rhesus embryonic stem cells to neural progenitors and neurons

Cited by 32 publications

References 41 publications

The Apical Polarity Determinant Crumbs 2 Is a Novel Regulator of ESC-Derived Neural Progenitors

The Apical Polarity Determinant Crumbs 2 Is a Novel Regulator of ESC-Derived Neural Progenitors

Establishment of Novel Embryonic Stem Cell Lines Derived from the Common Marmoset ( Callithrix jacchus )

The Differentiation of Neuronal Cells from Mouse Embryonic Stem Cells

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