Screening for mammalian neural genes via fluorescence-activated cell sorter purification of neural precursors from
            <i>Sox1</i>
            -
            <i>gfp</i>
            knock-in mice

Aubert, Jacques; Stavridis, Marios P.; Tweedie, Susan; O'Reilly, Michelle; Vierlinger, Klemens; Li, Meng; Ghazal, Peter; Pratt, Thomas; Mason, John O.; Roy, Douglas; Smith, Austin

doi:10.1073/pnas.1734197100

Cited by 231 publications

(203 citation statements)

References 47 publications

Supporting

Mentioning

196

Contrasting

Order By: Relevance

“…For size-defined oligosaccharides, degree of polymerization (dp) is dp2 for disaccharide, and so on. The 46C ES cell line was a gift from Austin Smith [30,31], and the Ext1 À/À line was generously provided by Prof. Dan Wells. PD173074 was used at 250 nM (Sigma Aldrich, St. Louis, www.sigmaaldrich.com).…”

Section: Methodsmentioning

confidence: 99%

Specific Glycosaminoglycans Modulate Neural Specification of Mouse Embryonic Stem Cells

et al. 2011

Self Cite

View full text Add to dashboard Cite

Mouse embryonic stem (mES) cells express a low sulfated form of heparan sulfate (HS). HS chains displayed by ES cells and their progeny become more complex and more sulfated during progression from pluripotency to neuroectodermal precursors. Sulfated epitopes are important for recognition and binding of a variety of ligands including members of the fibroblast growth factor (FGF) family. We demonstrated previously that mES cells lacking HS cannot undergo neural specification but this activity can be recovered by adding soluble heparin, a highly sulfated glycosaminoglycan (GAG). Therefore, we hypothesized that soluble GAGs might be used to support neural differentiation of HS competent cells and that the mechanisms underlying this activity might provide useful information about the signaling pathways critical for loss of pluripotency and early lineage commitment. In this study, we demonstrate that specific HS/heparin polysaccharides support formation of Sox1 1 neural progenitor cells from wild-type ES cells. This effect is dependent on sulfation pattern, concentration, and length of saccharide. Using a selective inhibitor of FGF signal transduction, we show that heparin modulates signaling events regulating exit from pluripotency and commitment to primitive ectoderm and subsequently neuroectoderm. Interestingly, we were also able to demonstrate that multiple receptor tyrosine kinases were influenced by HS in this system. This suggests roles for additional factors, possibly in cell proliferation or protection from apoptosis, during the process of neural specification. Therefore, we conclude that soluble GAGs or synthetic mimics could be considered as suitable low-cost factors for addition to ES cell differentiation regimes. STEM CELLS 2011;29:629-640 Disclosure of potential conflicts of interest is found at the end of this article.

show abstract

Section: Methodsmentioning

confidence: 99%

Specific Glycosaminoglycans Modulate Neural Specification of Mouse Embryonic Stem Cells

et al. 2011

Self Cite

View full text Add to dashboard Cite

show abstract

“…Sox1-GFP mouse ESCs 46 were cultured on gelatinized dishes in DMEM (GIBCO) supplemented with 15% FBS (GIBCO), 0.2 mM b-Mercaptoethanol, 1X MEM nonessential amino acids (GIBCO), 2 mM L-glutamine (GIBCO) and LIF. Embryoid bodies were formed by culturing ESCs in the absence of LIF.…”

Section: Methodsmentioning

confidence: 99%

A role for polyamine regulators in ESC self-renewal

Zhao

Goh

et al. 2012

Cell Cycle

View full text Add to dashboard Cite

E mbryonic stem cells (ESCs) dependon extensive regulatory networks to coordinate their self-renewal and differentiation. The polyamine pathway regulator AMD1 was recently implicated in ESC self-renewal and directed differentiation of ESCs to neural precursor cells (NPCs). The polyamines spermine and spermidine are essential for a wide range of biological processes, and their levels are tightly regulated. Here, we review the polyamine pathway and discuss how it can impact polyamine levels, cellular methylation and hypusinated EIF5A levels. We discuss how it could feed into regulation of ESC self-renewal and directed differentiation. We show that in addition to AMD1, a second rate-limiting enzyme in the polyamine pathway, ODC1, can also promote ESC self-renewal, and that both Amd1 and Odc1 can partially substitute for Myc during cellular reprogramming. We propose that both Amd1 and Odc1 are essential regulators of ESCs and function to ensure high polyamine levels to promote ESC self-renewal.

show abstract

“…To this end, we studied animals from the Sox1-GFP mouse line, known to provide a faithful GFP reporter signal indicative of Sox1 expression [10,11]. Cerebella from Sox1-GFP +/− mice were collected at stages ranging from birth (P0) to postnatal day 25 (P25), and the expression of the GFP reporter was analyzed in tissue sections (Figure 1).…”

Section: Developmental Distribution Of Sox1-expressing Cells In the Dmentioning

confidence: 99%

“…Since expression of this construct downstream of the endogenous Sox1 promoter [11] confers puromycin resistance to Sox1-expressing cells, the application of puromycin selection to cerebellar samples collected after enzymatic treatment provided an alternative to mechanical disruption in order to gradually break down the tissue, allow the progressive release of individual cells and promote the selective culture of Sox1-expressing cells.…”

Section: S2)mentioning

confidence: 99%

Dynamic distribution and stem cell characteristics of Sox1-expressing cells in the cerebellar cortex

Alcock

Sottile

2009

Cell Res

View full text Add to dashboard Cite

Bergmann glia cells are a discrete radial glia population surrounding Purkinje cells in the cerebellar cortex. Although Bergmann glia are essential for the development and correct arborization of Purkinje cells, little is known about the regulation of this cell population after the developmental phase. In an effort to characterize this population at the molecular level, we have analyzed marker expression and established that adult Bergmann glia express Sox1, Sox2 and Sox9, a feature otherwise associated with neural stem cells (NSCs). In the present study, we have further analyzed the developmental pattern of Sox1-expressing cells in the developing cerebellum. We report that before becoming restricted to the Purkinje cell layer, Sox1-positive cells are present throughout the immature tissue, and that these cells show characteristics of Bergmann glia progenitors. Our study shows that these progenitors express Sox1, Sox2 and Sox9, a signature maintained throughout cerebellar maturation into adulthood. When isolated in culture, the Sox1-expressing cerebellar population exhibited neurosphere-forming ability, NSC-marker characteristics, and demonstrated multipotency at the clonal level. Our results show that the Bergmann glia population expresses Sox1 during cerebellar development, and that these cells can be isolated and show stem cell characteristics in vitro, suggesting that they could hold a broader potential than previously thought.

show abstract

Screening for mammalian neural genes via fluorescence-activated cell sorter purification of neural precursors from Sox1 - gfp knock-in mice

Cited by 231 publications

References 47 publications

Specific Glycosaminoglycans Modulate Neural Specification of Mouse Embryonic Stem Cells

Specific Glycosaminoglycans Modulate Neural Specification of Mouse Embryonic Stem Cells

A role for polyamine regulators in ESC self-renewal

Dynamic distribution and stem cell characteristics of Sox1-expressing cells in the cerebellar cortex

Contact Info

Product

Resources

About