Journal: Cell Biochemistry and Function

Article Title: Morphological and Optical Characterization of NRAS‐Mutant Melanoma Cells and Primary Melanocytes via Quantitative Phase Imaging With Digital Holographic Microscopy

doi: 10.1002/cbf.70156

Figure Lengend Snippet: K‐Means Elbow Plot and Hierarchical Clustering Dendrogram comparing melanocytes and SK‐MEL‐2 cells using morphological, positional, geometric, phaseshift, roughness, and texture parameters. The K‐Means Elbow Plot (A) characterizes the optimal number of data clusters within the melanocyte and SK‐MEL‐2 data sets, as demonstrated by the “elbow” in the plot. The Hierarchical Clustering Dendrogram (B) visualizes the clustering of data points based on their similarity. The great height difference before breaking into two branches demonstrates a great difference between the two cell lines. These findings from the K‐Means Elbow Plot and Hierarchical Clustering Dendrograms suggest that the selected features highlight the differences between melanocytes and SK‐MEL‐2 cells. The plots were visualized using R (version 4.0.5). Analysis included 21,908 melanocyte measurements and 21,908 SK‐MEL‐2 measurements.

Article Snippet: Primary epidermal melanocytes (HEMa, ATCC PCS‐200‐013) were cultured in a medium consisting of Dermal Cell Basal Medium (ATCC PCS‐200‐030) and the Adult Melanocyte Growth Kit (ATCC PCS‐200‐042) at 37°C with 5% CO 2 .

Techniques:

Journal: Cell Biochemistry and Function

Article Title: Morphological and Optical Characterization of NRAS‐Mutant Melanoma Cells and Primary Melanocytes via Quantitative Phase Imaging With Digital Holographic Microscopy

doi: 10.1002/cbf.70156

Figure Lengend Snippet: Violin plots of morphological features for melanocytes versus SK‐MEL‐2 melanoma cells. The melanocytes exhibited smaller, less polarized distributions of (A) average areas, (B) boxed breadth, and (C) boxed length, suggesting smaller areas with narrower sets of dimensions than SK‐MEL‐2 cells. The SK‐MEL‐2 cells exhibited similar values to the melanocytes, however, in (D) eccentricity and (E) hull convexity, suggesting similar elongation with convex shapes. The melanocytes exhibited greater (F) irregular boundaries with lower (G) optical volumes, (H) perimeter lengths, and (I) shape convexities. Additionally, melanocytes exhibited lower, more concentrated (J) optical path length avg, (K) optical path length max, (L) optical thickness avg, and (M) optical thickness max, indicating smaller thicknesses and path lengths than the SK‐MEL‐2 cells. Melanocytes are represented in the color blue, and SK‐MEL‐2 cells are visualized in the color orange. The plots were generated in R (version 4.4.0), and the statistical significance was indicated as **** for p < 0.0001 comparing the two cell lines. Analysis included 21,908 melanocyte measurements and 21,908 SK‐MEL‐2 measurements.

Article Snippet: Primary epidermal melanocytes (HEMa, ATCC PCS‐200‐013) were cultured in a medium consisting of Dermal Cell Basal Medium (ATCC PCS‐200‐030) and the Adult Melanocyte Growth Kit (ATCC PCS‐200‐042) at 37°C with 5% CO 2 .

Techniques: Generated

Journal: Cell Biochemistry and Function

Article Title: Morphological and Optical Characterization of NRAS‐Mutant Melanoma Cells and Primary Melanocytes via Quantitative Phase Imaging With Digital Holographic Microscopy

doi: 10.1002/cbf.70156

Figure Lengend Snippet: Violin plots of positional and geometric parameters for melanocytes versus SK‐MEL‐2 cells. The SK‐MEL‐2 cells exhibited more distributed values across the (A) boxed center position X and (B) Y; (C) centroid position X and (D) Y; and (E) peak position X and (F) Y, suggesting greater variation in the location of the geometric center and center of mass. Melanocytes are represented in the color blue, and SK‐MEL‐2 cells are visualized in the color orange. The plots were generated in R (version 4.4.0), and the statistical significance was indicated as **** for p < 0.0001 comparing the two cell lines. Analysis included 21,908 melanocyte measurements and 21,908 SK‐MEL‐2 measurements.

Article Snippet: Primary epidermal melanocytes (HEMa, ATCC PCS‐200‐013) were cultured in a medium consisting of Dermal Cell Basal Medium (ATCC PCS‐200‐030) and the Adult Melanocyte Growth Kit (ATCC PCS‐200‐042) at 37°C with 5% CO 2 .

Techniques: Generated

Journal: Cell Biochemistry and Function

Article Title: Morphological and Optical Characterization of NRAS‐Mutant Melanoma Cells and Primary Melanocytes via Quantitative Phase Imaging With Digital Holographic Microscopy

doi: 10.1002/cbf.70156

Figure Lengend Snippet: Violin plots of phaseshift and roughness parameters for melanocytes versus SK‐MEL‐2 cells. The SK‐MEL‐2 cells exhibited greater, more distributed values for (A) phaseshift avg, (B) phaseshift std. dev, and (C) phaseshift sum, indicating greater phase shifts. The melanocytes exhibited greater (D) roughness avg, suggesting rougher surfaces compared to the SK‐MEL‐2 cells. Melanocytes are represented in the color blue, and SK‐MEL‐2 cells are visualized in the color orange. The plots were generated in R (version 4.4.0), and the statistical significance was indicated as **** for p < 0.0001 comparing the two cell lines. Analysis included 21,908 melanocyte measurements and 21,908 SK‐MEL‐2 measurements.

Article Snippet: Primary epidermal melanocytes (HEMa, ATCC PCS‐200‐013) were cultured in a medium consisting of Dermal Cell Basal Medium (ATCC PCS‐200‐030) and the Adult Melanocyte Growth Kit (ATCC PCS‐200‐042) at 37°C with 5% CO 2 .

Techniques: Generated

Journal: Cell Biochemistry and Function

Article Title: Morphological and Optical Characterization of NRAS‐Mutant Melanoma Cells and Primary Melanocytes via Quantitative Phase Imaging With Digital Holographic Microscopy

doi: 10.1002/cbf.70156

Figure Lengend Snippet: Violin plots of texture parameters for melanocytes versus SK‐MEL‐2 cells. The melanocytes exhibited greater, more concentrated values in (A) texture clustershade, but lower values in (B) texture clustertendency. The SK‐MEL‐2 cells exhibited lower (C) texture contrast but greater (D) texture correlation and (E) texture energy, suggesting greater uniformity in image texture. Additionally, the melanocytes exhibited greater (F) texture entropy but smaller (G) texture max. prob. and (H) texture homogeneity, suggesting more melanocyte nonuniformity and complexity. The plots were visualized in R (version 4.4.0), with melanocytes in blue and SK‐MEL‐2 cells in orange. Statistical significance was indicated as **** for p < 0.0001 comparing the two cell lines. Analysis included 21,908 melanocyte measurements and 21,908 SK‐MEL‐2 measurements.

Article Snippet: Primary epidermal melanocytes (HEMa, ATCC PCS‐200‐013) were cultured in a medium consisting of Dermal Cell Basal Medium (ATCC PCS‐200‐030) and the Adult Melanocyte Growth Kit (ATCC PCS‐200‐042) at 37°C with 5% CO 2 .

Techniques:

Journal: Cell Biochemistry and Function

Article Title: Morphological and Optical Characterization of NRAS‐Mutant Melanoma Cells and Primary Melanocytes via Quantitative Phase Imaging With Digital Holographic Microscopy

doi: 10.1002/cbf.70156

Figure Lengend Snippet: PCA and t‐SNE plots comparing melanocytes and SK‐MEL‐2 cells using morphological, positional, geometric, phaseshift, roughness, and texture parameters. The PCA plot (A) simplifies and separates the data across the first two principal components (PC1, PC2), visualizing variance while maintaining the global structure of the data. The t‐SNE plot (B) simplifies the high‐dimensional data and visualizes it across the two t‐SNE axes, maintaining the local structure of the data to a greater degree than the PCA plot. The clear separation between the melanocyte and SK‐MEL‐2 clusters in both the PCA and t‐SNE plots suggests significant differences between the two cell lines in terms of the parameters measured. The plots were generated in Python. Melanocytes are represented in the color blue, and SK‐MEL‐2 cells are visualized in the color orange. Analysis included 21,908 melanocyte measurements and 21,908 SK‐MEL‐2 measurements.

Article Snippet: Primary epidermal melanocytes (HEMa, ATCC PCS‐200‐013) were cultured in a medium consisting of Dermal Cell Basal Medium (ATCC PCS‐200‐030) and the Adult Melanocyte Growth Kit (ATCC PCS‐200‐042) at 37°C with 5% CO 2 .

Techniques: Generated

Journal: Cell Biochemistry and Function

Article Title: Morphological and Optical Characterization of NRAS‐Mutant Melanoma Cells and Primary Melanocytes via Quantitative Phase Imaging With Digital Holographic Microscopy

doi: 10.1002/cbf.70156

Figure Lengend Snippet: Ranked Feature Importance Diagram in differentiating melanocytes from SK‐MEL‐2 cells based on the Random Forest model. The Ranked Feature Importance Diagram reveals the parameters most significant in differentiating between the melanocytes and SK‐MEL‐2 cells. The optical parameters were demonstrated as the most significant, while the roughness parameters were the least significant. The diagram was generated using R (version 4.0.5). Analysis included 21,908 melanocyte measurements and 21,908 SK‐MEL‐2 measurements.

Article Snippet: Primary epidermal melanocytes (HEMa, ATCC PCS‐200‐013) were cultured in a medium consisting of Dermal Cell Basal Medium (ATCC PCS‐200‐030) and the Adult Melanocyte Growth Kit (ATCC PCS‐200‐042) at 37°C with 5% CO 2 .

Techniques: Generated

Journal: Cell Biochemistry and Function

Article Title: Morphological and Optical Characterization of NRAS‐Mutant Melanoma Cells and Primary Melanocytes via Quantitative Phase Imaging With Digital Holographic Microscopy

doi: 10.1002/cbf.70156

Figure Lengend Snippet: Mean area, volume, and diameter of melanocytes versus SK‐MEL‐2 cells over 48 h. Bar graphs highlighted the general increase in SK‐MEL‐2 (A) mean cell area, (B) mean cell volume, and (C) mean cell diameter at 0, 12, 24, 36, and 48 h, while the melanocytes remained relatively consistent. Melanocytes are represented in the color blue, and SK‐MEL‐2 cells are visualized in the color orange. The plots were generated in R (version 4.4.0). Error bars represent the standard deviation of the measurements, exhibiting variability and possible significance across the parameters measured. Analysis included 21,908 melanocyte measurements and 21,908 SK‐MEL‐2 measurements.

Article Snippet: Primary epidermal melanocytes (HEMa, ATCC PCS‐200‐013) were cultured in a medium consisting of Dermal Cell Basal Medium (ATCC PCS‐200‐030) and the Adult Melanocyte Growth Kit (ATCC PCS‐200‐042) at 37°C with 5% CO 2 .

Techniques: Generated, Standard Deviation

Journal: Scientific Reports

Article Title: A tool kit for rapid cloning and expression of recombinant antibodies

doi: 10.1038/srep05885

Figure Lengend Snippet: (A–B) Flow cytometric histograms showing fluorescent intensity of cells incubated with purified IgG/IgE antibodies plus secondary FITC-labelled goat anti-human IgG/IgE (open) and cells with secondary antibody only (filled) against the number of cells. (A) Flow cytometric assessment of anti-CSPG4 IgG 1 /IgE antibodies shows specific binding to native CSPG4 antigen present on the cell surface of A375 melanoma cells and no binding above background to primary melanocytes. (B) Fc regions of anti-CSPG4 IgG 1 and 102.1F10 IgG 4 isotypes demonstrate effector-binding to U937 monocytic cell line, expressing human Fcγ receptors. The IgE antibody isotypes also bind similarly to RBL SX38 mast cells, expressing human FcεR1 receptor. (C) Immunofluorescence staining of A375 cells confirms specific binding of anti-CSPG4 IgG 1 /IgE and no background binding with isotype control hapten specific NIP-IgG 1 /IgE detected by goat anti-human IgG/IgE-FITC. (D) Grass pollen allergen specificity is confirmed by anti-human sandwich ELISA, showing specific binding of recombinant 102.1F10 IgG 4 /IgE and original patient serum to the ELISA plate-bound Phl p 7 allergen and no binding above background with unspecific human myeloma IgG 4 and MOv18 IgE isotype controls.

Article Snippet: Primary human epidermal melanocytes (PCS-200-012, ATCC) were grownin Dermal Cell Basal Medium (PCS-200-030, ATCC) and supplemented with the Melanocyte Growth Kit (PCS-200-041, ATCC).

Techniques: Incubation, Purification, Binding Assay, Expressing, Immunofluorescence, Staining, Control, Sandwich ELISA, Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Diagnostics

Article Title: Alpha-Enolase (ENO1) Correlates with Invasiveness of Cutaneous Melanoma—An In Vitro and a Clinical Study

doi: 10.3390/diagnostics12020254

Figure Lengend Snippet: The expression level of ENO1 in the cell lysates from primary melanocytes and melanoma cell lines. Representative Western blots showing ENO1 and Akt 1/2/3 expression (for normalization) in protein lysates obtained from the primary human melanocytes (HEM) and indicated melanoma cell lines ( a ). Densitometric ENO1/Akt ratios are shown as mean values ( n = 3 except for HEM, n = 2) ± standard error of the mean (SEM) ( b ). The significance level was set at p = 0.001–0.0001 (***).

Article Snippet: Human epidermal melanocytes, adult (HEMa, 104−05A) and primary human epidermal melanocytes (lightly pigmented) (HEMn-LP, C0025C) were purchased from Cell Applications Inc (San Diego, CA, USA), and Cascade Biologics/Gibco (Carlsbad, CA, USA), respectively.

Techniques: Expressing, Western Blot

Journal: Experimental dermatology

Article Title: The nuclear factor (erythroid-derived 2)-like 2 (NRF2) antioxidant response promotes melanocyte viability and reduces toxicity of the vitiligo-inducing phenol monobenzone

doi: 10.1111/exd.13350

Figure Lengend Snippet: (a) The schema describes the process of ROS neutralization and highlights NRF2-targets, NQO1 and PRDX6 (in bold), that perform the same functions as generalized antioxidants. Total protein was extracted from four untreated cell lines derived from human skin. Cutaneous lines compared consisted of neonatal human epidermal melanocytes (NHEM), adult human epidermal melanocytes (AHEM), neonatal human dermal fibroblasts (NHDF) and neonatal human epidermal keratinocytes (NHEK). Protein levels of NRF2, SOD2, NQO1, PRDX6, and Actin (loading control) were measured by Western blot analysis. (b) NHEM were transfected with siRNAs against NRF2, NQO1, and PRDX6 and viability measured relative to non-target (NT) siRNA control. (c) NHDF and (d) NHEK were treated with siRNAs as in (b) in addition to t-BHP (250 μM) for 24 hours and viability measured relative to untreated NT siRNA control. Relative viability data are presented as mean ± SD, n=4 *** p < 0.001; **** p < 0.0001.

Article Snippet: Epidermal melanocytes from neonatal foreskin (NHEM, 3 lines established from normally pigmented, unrelated individuals) and an adult epidermal melanocyte (AHEM) line were purchased and cultured in DermaLife-M culture medium (Lifeline Cell Technology, Frederick, MD).

Techniques: Neutralization, Derivative Assay, Control, Western Blot, Transfection

Journal: Experimental dermatology

Article Title: The nuclear factor (erythroid-derived 2)-like 2 (NRF2) antioxidant response promotes melanocyte viability and reduces toxicity of the vitiligo-inducing phenol monobenzone

doi: 10.1111/exd.13350

Figure Lengend Snippet: (a) Relative viability of normal adult human epidermal melanocytes (AHEM) and melanocytes derived from perilesional (PLVM) and non-lesional (NLVM) skin from a vitiligo patient were compared after exposure to increasing concentrations of MBEH for 96 hours. (b) Relative viability of immortalized control melanocytes (PIG1) and immortalized vitiligo melanocytes (PIG3V) compared after exposure to increasing concentrations of MBEH for 96 hours. PIG1 and PIG3V were stimulated with MBEH (300 μM) for increasing periods up to 24 hours and mRNA expression of (c) HMOX1 and (d) GSH pathway members GCLC, GCLM, and GPX1 was measured. The relative viability data is presented as mean ± SD. mRNA expression data are presented as mean ± SEM, n=3. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Epidermal melanocytes from neonatal foreskin (NHEM, 3 lines established from normally pigmented, unrelated individuals) and an adult epidermal melanocyte (AHEM) line were purchased and cultured in DermaLife-M culture medium (Lifeline Cell Technology, Frederick, MD).

Techniques: Derivative Assay, Control, Expressing