human healthy skin fibroblast  (ATCC)

Journal: Scientific Reports

Article Title: Synthetic aporphine alkaloids are potential therapeutics for Leigh syndrome

doi: 10.1038/s41598-024-62445-w

Figure Lengend Snippet: ATP production capacity of LS patient-derived skin fibroblasts. No significant enhancement of ATP production upon administration of apomorphine, D31, D55, or D40.

Article Snippet: Healthy skin fibroblasts purchased from Promo Cell GmbH (#C-12300; Heidelberg, Germany) were used as controls.

Techniques: Derivative Assay

Journal: Scientific Reports

Article Title: Synthetic aporphine alkaloids are potential therapeutics for Leigh syndrome

doi: 10.1038/s41598-024-62445-w

Figure Lengend Snippet: GDF-15 suppression in RSL3 loaded LS fibroblasts of D31, D55, and D40. Compared to the RSL3-only, the groups co-administered with apomorphine, ferrostatin-1, D31, D55, and D40 showed a significant decrease in the concentration of GDF-15 in cell supernatant of LS patient-derived skin fibroblasts.

Article Snippet: Healthy skin fibroblasts purchased from Promo Cell GmbH (#C-12300; Heidelberg, Germany) were used as controls.

Techniques: Concentration Assay, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Generation and Characterization of Human iPSC-Derived Astrocytes with Potential for Modeling X-Linked Adrenoleukodystrophy Phenotypes

doi: 10.3390/ijms26041576

Figure Lengend Snippet: Generation of iPSCs from patient-derived skin fibroblasts. ( A ) Representative phase contrast images of CTL, AMN, and cALD iPSCs. ( B ) Representative alkaline-phosphatase-stained images of CTL, AMN, and cALD iPSCs. ( C ) Immunostaining for pluripotency markers: NANOG, TRA1–60, SOX2, and SSEA. Nuclei are stained with DAPI (blue). ( D ) RT-qPCR quantification of pluripotency markers (ratio against L27) in CTL, AMN iPSCs ( n = 3). ( E ) Representative immunocytochemistry showed iPSC-derived cells positive for markers of three germ layers following differentiation: endoderm (SOX17), mesoderm (BRACHYURY and DESMIN), ectoderm (OXT2 and TUBB3). AMN: Adrenomyeloneuropathy; cALD: Cerebral adrenoleukodystrophy; CTL: Control; iPSC: Induced pluripotent stem cell. Scale: 100 µM ( A , B ) and 50 µM ( C , E ).

Article Snippet: Healthy human skin CTL fibroblasts (GM08402; 32-year-old male, GM03348; 10-year-old male), AMN fibroblasts (GM17819; 32-year-old male, GM07675; 22-year-old male), and cALD fibroblasts (GM04904; 11-year-old male, GM04496; 6-year-old male) were obtained from the National Institute for General Medical Sciences human genetic cell repository at Coriell Institute for Medical Research, Camden, NJ, USA.

Techniques: Derivative Assay, Staining, Immunostaining, Quantitative RT-PCR, Immunocytochemistry, Control

Journal: Advanced Science

Article Title: Targeting Nuclear Mechanics Mitigates the Fibroblast Invasiveness in Pathological Dermal Scars Induced by Matrix Stiffening

doi: 10.1002/advs.202308253

Figure Lengend Snippet: In human keloid tissues, activation of dermal fibroblasts is associated with an abnormal increase in matrix stiffness. a) Masson‘s trichrome staining of human keloid tissues shows heterogeneity in architecture, with regions resembling normal skin tissues and regions with abnormally thick collagen bundles (blue, * ). These regions were labeled as “Soft” and “Stiff”, respectively. Scale bar, 2 mm (0.2 ×), 40 µm (10 ×), and 10 µm (40 ×). b,c) Stiffness scanning of human keloid tissues (45 locations with a spacing of 300 µm horizontally and 500 µm vertically in an area of 0.9 mm width × 0.75 mm length) showing a spatial profile of tissue stiffness (b) and heterogeneity in stiffness ranging from 0.5 kPa to 184.2 kPa ( n = 100 locations) (c). d) Immunohistochemical staining for α‐SMA (brown) in keloid tissue. Scale bar, 40 µm (10 ×), and 10 µm (40 ×). e) YAP staining (left, brown) and quantification of nuclear YAP intensity (right, n = 100 cells/condition from 3 independent donors). Scale bar, 40 µm (10 ×), and 10 µm (40 ×). Keloidal thick collagen bundles are indicated by asterisks. f) Stiffness of keloid‐mimicking PDMS substrates ( n = 3). g) α‐SMA staining (left, green) and quantification (right, n = 20 fields/condition from 3 independent experiments) showing increased α‐SMA + cells (%) in keloid fibroblasts cultured on stiff substrates, similar to the in vivo observation. Nuclei are stained with DAPI (blue). Scale bar, 100 µm. **** p < 0.0001; two‐tailed paired Student's t ‐test.

Article Snippet: Primary human dermal fibroblasts isolated from healthy skin or keloids were purchased from Fisher Scientific or ATCC, respectively (See Table , Supporting Information, for detailed information on cells).

Techniques: Activation Assay, Staining, Labeling, Immunohistochemical staining, Cell Culture, In Vivo, Two Tailed Test

Journal: Advanced Science

Article Title: Targeting Nuclear Mechanics Mitigates the Fibroblast Invasiveness in Pathological Dermal Scars Induced by Matrix Stiffening

doi: 10.1002/advs.202308253

Figure Lengend Snippet: Keloid fibroblasts exhibit elevated mechano‐sensitivity independent of biochemical cues compared to normal fibroblasts. Normal fibroblasts and keloid fibroblasts were stained for paxillin, F‐actin, YAP, and MRTF‐A after 2 days of culture on either soft or stiff substrates in the absence of TGF‐β1. a,b) analysis of focal adhesion (FA) formations. Immunostaining for paxillin (green) (a) and quantification of the number ( n = 30 FA/cell), size ( n = 100 FA/condition), and aspect ratio (AR, n = 100 FA/condition) of focal adhesions (b). Nuclei are stained with DAPI (blue). Scale bar, 50 µm (main images), and 15 µm (inserts). c,d) Actin development analysis. Representative images of F‐actin staining (red) on the basal and apical planes (c) and quantitative analysis of the F‐actin anisotropy, the number, and thickness of F‐actin bundles (d, n = 50–200 cells/condition). Scale bar, 25 µm (main images), and 10 µm (insets). e) Representative images of immunostaining showing intracellular localization of YAP (top, green) or MRTF‐A (bottom, green). Scale bar, 40 µm. f) Quantitative analysis of nuclear YAP or MRTF‐A positive cells ( n = 10–20 fields/condition). Data are representative of at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; two‐way ANOVA followed by Tukey's post‐hoc tests.

Article Snippet: Primary human dermal fibroblasts isolated from healthy skin or keloids were purchased from Fisher Scientific or ATCC, respectively (See Table , Supporting Information, for detailed information on cells).

Techniques: Staining, Immunostaining

Journal: Advanced Science

Article Title: Targeting Nuclear Mechanics Mitigates the Fibroblast Invasiveness in Pathological Dermal Scars Induced by Matrix Stiffening

doi: 10.1002/advs.202308253

Figure Lengend Snippet: Matrix stiffness triggers changes in the nuclear shape of keloid fibroblasts, which are then accompanied by chromatin reorganization and histone modification. a,b) Nuclear morphology analysis showing color‐coded images of 3D nuclear morphology (a) along with quantitative analysis of nuclear flatting index (b, n = 50–56 nuclei/condition). c,d) Representative color‐coded images of DAPI‐stained nuclei (c) subjected to chromatin condensation analysis (d, n = 200 nuclei/condition). e–h) Immunostaining for histone 3 acetylation (e, n = 200 nuclei/condition) and H3K9 methylation (g, n = 100 nuclei/condition), and quantification of mean intensity of the immunostaining (f and h). Scale bar, 10 µm (c, e, and g). i) Gene expression of HDAC 1, 11, and HAT 1 ( n = 4/condition). Data represent mean ± s.d. of n and are representative of at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; two‐way ANOVA followed by Tukey's post‐hoc tests.

Article Snippet: Primary human dermal fibroblasts isolated from healthy skin or keloids were purchased from Fisher Scientific or ATCC, respectively (See Table , Supporting Information, for detailed information on cells).

Techniques: Modification, Staining, Immunostaining, Methylation, Gene Expression

Journal: Advanced Science

Article Title: Targeting Nuclear Mechanics Mitigates the Fibroblast Invasiveness in Pathological Dermal Scars Induced by Matrix Stiffening

doi: 10.1002/advs.202308253

Figure Lengend Snippet: Alteration in the nuclear shape of keloid fibroblasts is associated with modifications in the nuclear lamina. a,b) Lamin A/C expression in NFs and KFs after culture on either soft or stiff substrates for 2 and 7 days. Representative western blots (a) and quantification (b, n = 3–6 replicates/condition). c) Representative images of immunostaining for lamin A/C showing its distribution within the nucleus. XZ‐ and YZ‐axes projections on the equatorial plane (EP) of the nucleus (top). Maximum intensity projection (MIP) of a series of z‐stacks images covering the entire nuclear region (middle), along with line scans of fluorescence intensity (F. I.) from the immunostaining images (bottom). Scale bar, 10 µm. Dotted lines mark the edges of the nucleus. d–g) Lamin A/C expression in human keloid tissues. Representative images of immunohistochemical staining for lamin A/C in human keloid tissues (d, left). Scale bar, 2 mm (0.2 ×), 40 µm (10 ×), and 10 µm (40 ×). Line scans showing the intensity profiles of lamin A/C staining in the nucleus (d, right). Arrows mark the edges of the nucleus. The lamin A/C intensity (e), nuclear area (f, left), and contour ratio (f, right) of nuclei quantified based on the immunostaining ( n = 698 cells/condition from three independent donors). Correlation plots show a negative correlation between lamin A/C intensity and contour ratio in both soft and stiff regions (g). The correlation coefficient (Pearson's r ) was determined by linear fits as in (g, dash line for Soft, and solid line for Stiff). h) Representative images of co‐immunostaining for lamin A/C (green) and a‐SMA (red) in human keloid tissues. Scale bar, 10 µm (main) and 50 µm (insert). i) Cluster map displaying the correlation of lamin A/C and α‐SMA intensity in soft (green) and stiff (red) regions ( n = 50–54 cells/condition from three independent donors). Data represent mean ± s.d. of n and are representative of at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; one‐way ANOVA followed by Tukey's post‐hoc tests.

Article Snippet: Primary human dermal fibroblasts isolated from healthy skin or keloids were purchased from Fisher Scientific or ATCC, respectively (See Table , Supporting Information, for detailed information on cells).

Techniques: Expressing, Western Blot, Immunostaining, Fluorescence, Immunohistochemical staining, Staining

Journal: Advanced Science

Article Title: Targeting Nuclear Mechanics Mitigates the Fibroblast Invasiveness in Pathological Dermal Scars Induced by Matrix Stiffening

doi: 10.1002/advs.202308253

Figure Lengend Snippet: Nuclear softening, along with weak adhesion, accelerates confined migration in keloid fibroblasts. a–d) Nuclear deformability analysis. Representative time‐lapse image series of the deformation of fluorescently labeled cell nuclei in the microfluidic micropipette aspiration device (left) with a schematic (top right) (a). Scale bar, 10 µm. NFs, KFs, or KFs transfected with mCherry‐LMNA plasmid (OE) were pre‐cultured on either soft or stiff substrates for 2 days before the assay. The relative proportions of populations depending on nuclear deformability (b). The nuclear deformation rate was measured based on the protrusion length change over time upon aspiration (c, n > 30 nuclei/condition). Nuclear protrusion profiles for 180 sec of aspiration (d, n = 22–45 nuclei/condition). The slope ( m ) was determined by linear fits as shown in (d). e,f) Transwell migration assays on cells pre‐cultured under each condition for 2 days. An FE‐SEM image of transwell with (8 µm‐diameter pores) (top left), and pseudocolor images of crystal violet staining (rainbow) following the transwell migration assay (right) (e) and quantification (f, n = 10 field/condition). Scale bar, 200 µm. g,h) Analysis of focal adhesion formations in response to TGF‐β1. Representative co‐staining images of paxillin (green) and F‐actin (red) (g). Scale bar, 50 µm (main images), and 15 µm (inserts). Quantitative analysis of the number ( n = 30 cells/condition), size ( n = 100 FA/condition), and aspect ratios ( n = 100 FA/condition) of focal adhesions based on the co‐staining images (h). i–k) 3D confined migration assays through polymeric dense fibrous matrices. Schematic of the assay, along with an FE‐SEM image showing the fibrous structure of the matrix with 3.7 µm‐diameter pores (i). After 2 days of pre‐culture on stiff substrates with TGF‐β1, cells were replated onto the fibrous matrices for the confined migration assay. 3D confocal reconstructions of cell (red) migration through fibrous networks (green) along with cross‐sectional views (j). Nuclei are stained with DAPI (blue). Scale bar, 100 µm. Quantification of the percentage of infiltrated cells ( n = 11 fields/condition), cell infiltration depth ( n = 55 cells/condition from 10 independent experiments), and cell spreading area ( n = 55 cells/condition from 10 independent experiments) (k). Data are representative of at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; one‐way ANOVA followed by Tukey's post hoc tests for (b), and two‐tailed paired Student's t ‐test for (f), (h), and (k).

Article Snippet: Primary human dermal fibroblasts isolated from healthy skin or keloids were purchased from Fisher Scientific or ATCC, respectively (See Table , Supporting Information, for detailed information on cells).

Techniques: Migration, Labeling, Transfection, Plasmid Preparation, Cell Culture, Staining, Transwell Migration Assay, Two Tailed Test

Journal: Advanced Science

Article Title: Targeting Nuclear Mechanics Mitigates the Fibroblast Invasiveness in Pathological Dermal Scars Induced by Matrix Stiffening

doi: 10.1002/advs.202308253

Figure Lengend Snippet: Preventing lamin A/C‐driven nuclear softening blunts the aggressive confined migration behavior of keloid fibroblasts. a) Representative images of immunostaining for lamin A/C (red) in wild type (WT) or lamin A/C‐overexpressed (OE) keloid fibroblasts cultured on stiff substrates in the presence of TGF‐β1 for 2 days. XZ‐ and YZ‐axes projections on the equatorial plane of the nucleus. Scale bar, 10 µm. b) Line scans show the increased lamin A/C intensity and perinuclear accumulation in OE cells compared to WT cells. c) Representative western blots (left) and quantification of lamin A/C expression (right, n = 3 replicates/condition). d) Nuclear deformation rates evaluated by the microfluidic micropipette aspiration assay ( n > 35 cells/condition from 4 independent experiments). e) Representative images of crystal violet staining (purple) following the transwell (8 µm‐diameter pores) migration assay (left) and quantification (right, n = 10 fields/condition). Scale bar, 200 µm. f) 3D confocal reconstructions of cell (red) migration through polymeric fibrous networks (green) along with cross‐sectional views. Nuclei are stained with DAPI (blue). Scale bar, 100 µm. g) Quantification of the percentage of infiltrated cells ( n = 11 fields/condition), cell infiltration depth ( n = 55 cells from 10 independent experiments), and cell spreading area ( n = 55 cells from 10 independent experiments). h–l) Indirect regulation of lamin A/C by inhibiting force transmission to the nucleus through the actin cytoskeleton. Timeline showing that blebbistatin (Blebb) or latrunculin A (Lat‐A) were treated to KFs when KFs were seeded on stiff substrates (h). Representative western blots (i) and quantification of lamin A/C in KFs treated with either Blebb or Lat‐A (j, n = 3 replicates/condition). Representative images of crystal violet staining (purple) following the transwell (8 µm‐diameter pores) migration assay (k) and quantification (l, n = 10 fields/condition). Scale bar, 200 µm. Data represent mean ± s.d. of n and are representative of at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; two‐tailed paired Student's t ‐test or one‐way ANOVA followed by Tukey's post hoc tests or two‐way ANOVA followed by Tukey's post‐hoc tests.

Article Snippet: Primary human dermal fibroblasts isolated from healthy skin or keloids were purchased from Fisher Scientific or ATCC, respectively (See Table , Supporting Information, for detailed information on cells).

Techniques: Migration, Immunostaining, Cell Culture, Western Blot, Expressing, Staining, Transmission Assay, Two Tailed Test

Journal: Advanced Science

Article Title: Targeting Nuclear Mechanics Mitigates the Fibroblast Invasiveness in Pathological Dermal Scars Induced by Matrix Stiffening

doi: 10.1002/advs.202308253

Figure Lengend Snippet: A summary of how matrix stiffness modulates the mechano‐activation and invasive migration of keloid fibroblasts in confining environments. In keloid tissues, keloid fibroblasts (KFs) produce keloidal collagens, which form thickened, disorganized, and hyalinized collagen bundles. These collagens contribute to tissue stiffening. Additionally, keloidal tissues become even stiffer as skin tension intensifies, leading to abnormal mechanical forces exerted on cells. KFs display higher mechanosensitivity to matrix stiffness compared to normal fibroblasts (NFs). Upon TGF‐β1 stimulation, KFs exhibit distinct mechanoresponses, including reduced focal adhesion formation and enhanced actin development. Furthermore, nuclear deformability increases significantly, accompanied by decreased lamin A/C expression, increased euchromatin formation and histone acetylation, and disrupted anchoring of lamina‐associated chromatin. These events collectively facilitate the confined migration of KFs. Importantly, preventing lamin A/C‐driven nuclear softening through LMNA overexpression or actin disruption blunts the invasive migration of KFs.

Article Snippet: Primary human dermal fibroblasts isolated from healthy skin or keloids were purchased from Fisher Scientific or ATCC, respectively (See Table , Supporting Information, for detailed information on cells).

Techniques: Activation Assay, Migration, Expressing, Over Expression, Disruption

Journal: Saudi Pharmaceutical Journal : SPJ

Article Title: Phytochemical profiling of Salsola tetragona Delile by LC-HR/MS and investigation of the antioxidant, anti-inflammatory, cytotoxic, antibacterial and anti-SARS-CoV-2 activities

doi: 10.1016/j.jsps.2023.101731

Figure Lengend Snippet: IC 50 values of S. tetragona extracts estimated in the MTT cytotoxicity assay.

Article Snippet: Human healthy skin fibroblasts CCD-1079Sk (CRL-2097TM) and human breast cancer cells MCF7 (HTB-22TM) were acquired from American Type Culture Collection (ATCC, VA, USA).

Techniques: