neural stem cell markers nestin  (R&D Systems)

Journal: Reproductive Toxicology (Elmsford, N.y.)

Article Title: Exposure to human relevant mixtures of halogenated persistent organic pollutants (POPs) alters neurodevelopmental processes in human neural stem cells undergoing differentiation

doi: 10.1016/j.reprotox.2020.12.013

Figure Lengend Snippet: Characterization of hiPSC-derived neural stem cells (NSCs) undergoing differentiation into neurons and astrocytes in control culture. (A) Quantification of nestin, β-III-tubulin, MAP2 and GFAP positive cells shown as percentage of DAPI stained cells, comparing cells at 7, 14, 21 and 28 DIV of differentiation, to NSCs (1 DIV). (B) Quantification of Ki67+ proliferating cells undergoing differentiation as described in A. (C) Quantification of total BDNF protein levels, neurite length (β-III-tubulin staining) and the number of synapses (i.e., number of overlapping SYP/PSD95 spots in MAP2+ cells) (data in C graph were taken from and presented here in a different format). (D) Gene expression analysis of AhR gene in NSCs undergoing differentiation; AhR gene expression was normalized to Bactin and GAPDH and calibrated on NSCs at 1 DIV (mean ± S.E.M. of 3 biological replicates). (E) Representative immunocytochemical images of neuronal cells and astrocytes after 28 DIV; cells were stained for β-III-tubulin (red), GFAP (green), MAP2 (red), Ki67 (green), BDNF (green), PSD95 (red) and SYP (green). (F) Representative pictures of cells at 28 DIV, stained with MAP2, synaptophysin (SYP) and PSD95 antibodies, along with DAPI to identify nuclei and identified by masks using the Thermo Scientific HCS Studio Software (Cellomics platform). Values in A, B and C are normalised to undifferentiated NSCs (1 DIV), and are shown as mean ± S.E.M. of at least 4 biological replicates (* p < 0.05, ** p < 0.01, *** p < 0.001). (For interpretation of the references to colour in the Figure, the reader is referred to the web version of this article).

Article Snippet: Cells were also stained for glial fibrillary acidic protein (GFAP, chicken, 1:500, Abcam), the neural stem cell marker nestin (mouse, 1:200, Thermofisher) and the cell cycle marker Ki67 (rabbit, 1:1000, Abcam).

Techniques: Derivative Assay, Control, Staining, Gene Expression, Software

Journal: Reproductive Toxicology (Elmsford, N.y.)

Article Title: Exposure to human relevant mixtures of halogenated persistent organic pollutants (POPs) alters neurodevelopmental processes in human neural stem cells undergoing differentiation

doi: 10.1016/j.reprotox.2020.12.013

Figure Lengend Snippet: Effects of an acute treatment (3 days) with POP mixture containing PerF + Br + Cl on cell viability, proliferation and proportions of cell populations. (A) Starting on differentiation day 1 (1 DIV), hiPSC-derived NSCs were treated for 3 days with PerF + Br + Cl mixture at the concentrations 0.5, 1, 10, 100, 500 and 1000x (see ), and analysis of mitochondrial activity by means of resazurin test was performed (B). (C) Quantification of Ki67+ proliferating cells, and (D) nestin, MAP2 and GFAP positive cell percentages upon treatment with PerF + Br + Cl mixture at the concentrations 0.5, 1, and 1000 × . (E) Representative immunocytochemical images of NSCs (nestin+, green), and NSCs undergoing differentiation toward neurons (MAP2+, red) and astrocytes (GFAP+, green) in solvent control culture (upper panels) and upon 3 day treatment with PerF + Br + Cl mixture at 1000x concentration (10x magnification images). Values in B, C and D are normalised to solvent control cells and are shown as mean ± S.E.M. of 4 biological replicates (* p < 0.05, ** p < 0.01, *** p < 0.001). (For interpretation of the references to colour in the Figure, the reader is referred to the web version of this article).

Article Snippet: Cells were also stained for glial fibrillary acidic protein (GFAP, chicken, 1:500, Abcam), the neural stem cell marker nestin (mouse, 1:200, Thermofisher) and the cell cycle marker Ki67 (rabbit, 1:1000, Abcam).

Techniques: Derivative Assay, Activity Assay, Solvent, Control, Concentration Assay

Journal: Reproductive Toxicology (Elmsford, N.y.)

Article Title: Exposure to human relevant mixtures of halogenated persistent organic pollutants (POPs) alters neurodevelopmental processes in human neural stem cells undergoing differentiation

doi: 10.1016/j.reprotox.2020.12.013

Figure Lengend Snippet: Effects of a repeated dose exposure (14d and 28d) to different types of POP mixtures on proportion of diverse cell populations. (A) Quantification of nestin+, nestin+/Ki67+, MAP2+ and GFAP + cell percentages upon 14 or 28 day treatment with POP mixtures at the concentrations 0.5, 1, and 1000 × . (B) Representative immunocytochemical images of nestin (red) and Ki67 (green) (upper panels), and MAP2 (red) staining (lower panels) in solvent control culture and upon 28 day treatment with PerF only, Br only and Cl only mixtures at 1x concentration. Data are normalized to solvent control (0.1 % DMSO) and presented as mean ± S.E.M. with asterisks indicating statistical significance compared to solvent control (* p < 0.05, ** p < 0.01, *** p < 0.001). (For interpretation of the references to colour in the Figure, the reader is referred to the web version of this article).

Article Snippet: Cells were also stained for glial fibrillary acidic protein (GFAP, chicken, 1:500, Abcam), the neural stem cell marker nestin (mouse, 1:200, Thermofisher) and the cell cycle marker Ki67 (rabbit, 1:1000, Abcam).

Techniques: Staining, Solvent, Control, Concentration Assay

Journal: Stem Cells Translational Medicine

Article Title: Transplantation of Olfactory Stem Cells with Biodegradable Hydrogel Accelerates Facial Nerve Regeneration After Crush Injury

doi: 10.1002/sctm.15-0399

Figure Lengend Snippet: List of primers used for reverse transcriptase‐polymerase chain reaction

Article Snippet: Our results show that the neural stem cell markers Nestin and Musashi‐1 and the undifferentiated cell markers Sox‐2 and Nanog are expressed at the mRNA level in the olfactory spheres (Fig. B).

Techniques: Sequencing

Journal: Stem Cells Translational Medicine

Article Title: Transplantation of Olfactory Stem Cells with Biodegradable Hydrogel Accelerates Facial Nerve Regeneration After Crush Injury

doi: 10.1002/sctm.15-0399

Figure Lengend Snippet: Olfactory spheres and olfactory stem cells express neural stem cell markers. (A): Representative image of the floating olfactory spheres observed under a phase‐contrast microscope. (B): Reverse Transcriptase‐Polymerase Chain Reaction analysis of nestin, Musashi‐1, Sox‐2, and Nanog mRNA expression in the olfactory spheres. (C): Representative immunostaining images showing the protein expression of the neural stem cell markers Musashi‐1 ( Ca ), nestin ( Cb ), and OMP ( Cf ) as well as the expression of GFAP ( Ce ), βIII‐tubulin ( Ci ), and GalC ( Cj ), which label differentiated astrocytes, neurons, and oligodendrocytes, respectively. The nuclei were counterstained with Hoechst 33258 ( Cc , Cg , and Ck ), and merged images are shown in ( Cd ), ( Ch ), and ( Cl ). The scale bars indicate 100 μm ( Ca ) and 50 μm ( Cc ).

Article Snippet: Our results show that the neural stem cell markers Nestin and Musashi‐1 and the undifferentiated cell markers Sox‐2 and Nanog are expressed at the mRNA level in the olfactory spheres (Fig. B).

Techniques: Microscopy, Reverse Transcription, Polymerase Chain Reaction, Expressing, Immunostaining

Journal: Stem Cells Translational Medicine

Article Title: Transplantation of Olfactory Stem Cells with Biodegradable Hydrogel Accelerates Facial Nerve Regeneration After Crush Injury

doi: 10.1002/sctm.15-0399

Figure Lengend Snippet: Neural stem cell markers and neuronal markers in differentiated cells from the olfactory spheres. Representative immunostaining images showing the expression of the neural stem cell markers Musashi‐1 (A) , nestin (B) , and OMP ( F ) as well as GFAP ( E ), βIII‐tubulin (I) , and GalC (J) , which label astrocytes, neurons, and oligodendrocytes, respectively, in the differentiated olfactory stem cells. The nuclei were counterstained with Hoechst 33258 ( C , G , and K ), and merged images are shown in ( D ), ( H ), and (L) . The scale bars indicate 20 μm.

Article Snippet: Our results show that the neural stem cell markers Nestin and Musashi‐1 and the undifferentiated cell markers Sox‐2 and Nanog are expressed at the mRNA level in the olfactory spheres (Fig. B).

Techniques: Immunostaining, Expressing