Journal: International Journal of Molecular Sciences

Article Title: A Simple Ionic-Gelation Method for Chitosan Nanoparticle Synthesis and Standardized Protocols for Biological Safety Assessment: Antibacterial Activity, Phytotoxicity, and Biocompatibility

doi: 10.3390/ijms27083673

Figure Lengend Snippet: Cell viability of HDF cells treated with Ch NPs at concentrations of 2.5–160 µg/mL for 24 h. Viability was determined using the MTT assay and expressed as mean ± SE (n = 3). Different letters indicate statistically significant differences ( p < 0.05).

Article Snippet: The biocompatibility of Ch NPs was evaluated using primary HDF (ATCC ® PCS-201-012TM [ ], American Type Culture Collection, Manassas, VA, USA) and HaCaT cells (Cat. No. T0020001, AddexBio, San Diego, CA, USA).

Techniques: MTT Assay

Journal: International Journal of Molecular Sciences

Article Title: Hypothermically Stored Amnion Is Robust and Provides a Scaffold for Supporting Wound Healing by Retaining the Characteristics of Native Tissue

doi: 10.3390/ijms251910347

Figure Lengend Snippet: Fibroblast attachment and proliferation following seeding with human dermal fibroblasts (NHDF) and mouse fibroblasts (L929) onto hypothermically stored amniotic membranes. Cell number from AlamarBlue assessment showing metabolic activity of NHDFs ( A ) and L929s ( B ). ( C ) Representative immunofluorescence staining of seeded and non-seeded scaffolds at day 3 and 14; 40× magnification. Average ± standard deviation reported; ns denotes not significant; * denotes p < 0.05; ** denotes p < 0.01; *** denotes p < 0.001; **** denotes p < 0.0001 compared to non-seeded controls. Blue: nuclei; green: TGF-β1; red: f-actin (phalloidin); arrow: epithelial layer; S: spongy layer; Scale bar: 25 µm.

Article Snippet: Primary normal adult human dermal fibroblasts (NHDFs, lot 22TL073035 [female, age 46, Black], Lonza Biosciences, Walkersville, MD, USA) and two lots of primary mouse fibroblasts (L929, lots 14A015/70058860, ATCC, Bethesda, MD, USA) were thawed and cultured as per the manufacturer’s recommendations.

Techniques: Activity Assay, Immunofluorescence, Staining, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Hypothermically Stored Amnion Is Robust and Provides a Scaffold for Supporting Wound Healing by Retaining the Characteristics of Native Tissue

doi: 10.3390/ijms251910347

Figure Lengend Snippet: Structural characterization of fibroblasts seeded onto hypothermically stored amniotic membranes. Representative scanning electron microscopy images of non-seeded and human fibroblasts seeded ( A ) and mouse fibroblast seeded ( B ) grafts. For SEM, representative images are at 100× (scale bar: 500 µm) and 500× (scale bar: 100 µm).

Article Snippet: Primary normal adult human dermal fibroblasts (NHDFs, lot 22TL073035 [female, age 46, Black], Lonza Biosciences, Walkersville, MD, USA) and two lots of primary mouse fibroblasts (L929, lots 14A015/70058860, ATCC, Bethesda, MD, USA) were thawed and cultured as per the manufacturer’s recommendations.

Techniques: Electron Microscopy

Journal: Cells

Article Title: Impact of Progerin Expression on Adipogenesis in Hutchinson—Gilford Progeria Skin-Derived Precursor Cells

doi: 10.3390/cells10071598

Figure Lengend Snippet: Isolation of SKPs from control and HGPS fibroblasts. ( a ) Panel showing the protocol for SKP isolation. Briefly, fibroblasts were pelleted and treated with HBSS buffer (pH 5.7) for 30 min at 37 °C. Cells were cultured in SKP media containing DMEM low glucose, EGF, FGF, and B27. The flasks were agitated daily, and the spheroids were harvested at day 4 for analysis. ( b ) SKP formation from both control (GMO1651c, GMO5565) and HGPS (HGADFN127, HGADFN003) fibroblasts with 5 and 30% senescence (SNS). ( c , d ) Quantification of the number and the diameter of the spheroids from control and HGPS fibroblast cultures with 5 and 30% SNS at day 4. Values are presented as mean ± SD ( n = 3), not significant (ns), * p < 0.05, ( c , d ) unpaired t -test. HBSS: Hank’s Balanced Salt Solution, DMEM: Dulbecco′s modified Eagle medium, EGF: epidermal growth factor, FGF: fibroblast growth factor, SKPs: skin-derived precursor cells, SNS: senescence.

Article Snippet: The human primary dermal fibroblast cell lines, GMO5565 (3-year-old male), GMO1651 (13-year-old female) and GMO5567A (12-year-old male) were all obtained from the Coriell Institute for Medical Research (Camden, NJ, USA).

Techniques: Isolation, Control, Cell Culture, Modification, Derivative Assay

Journal: bioRxiv

Article Title: Altered DNA repair pathway engagement by engineered CRISPR-Cas9 nucleases

doi: 10.1101/2022.03.10.483793

Figure Lengend Snippet: a , Design of a gRNA and MDR template to introduce a precise edit. b , Precise editing and indel frequencies using MDR templates in dividing HEK293T cells. c , Precise editing and indel frequencies using MDR templates in non-dividing (quiescent) primary human dermal fibroblasts. * indicates p < 0.05 for precise editing frequency compared to wild-type Cas9. Data were analyzed by deep next-generation sequencing and represent means of n = 3 independent replicates with standard errors.

Article Snippet: HEK293T human embryonic kidney, HeLa human cervical cancer, A549 human lung cancer, Panc1 human pancreatic cancer, and primary human dermal fibroblasts from neonates (GlobalStem) were maintained in Dulbecco’s Modified Eagle’s Medium with high glucose, sodium pyruvate, and GlutaMAX (DMEM; ThermoFisher, 10569) supplemented with 10% Fetal Bovine Serum (FBS; ThermoFisher, 10438), and 100 U/mL Penicillin-Streptomycin (ThermoFisher, 15140).

Techniques: Introduce, Next-Generation Sequencing

Journal: bioRxiv

Article Title: Altered DNA repair pathway engagement by engineered CRISPR-Cas9 nucleases

doi: 10.1101/2022.03.10.483793

Figure Lengend Snippet: a , Cell cycle profiling of dividing and quiescent primary human dermal fibroblasts. b , Flow cytometry plots showing EdU-high (indicating S phase), Propidium Iodide-high (indicating G2 phase), and EdU-low Propidium Iodide-low (indicating G1 or G0 phase) cell population fractions. All data were analyzed by flow cytometry and represent means of n = 3 independent replicates with standard errors.

Article Snippet: HEK293T human embryonic kidney, HeLa human cervical cancer, A549 human lung cancer, Panc1 human pancreatic cancer, and primary human dermal fibroblasts from neonates (GlobalStem) were maintained in Dulbecco’s Modified Eagle’s Medium with high glucose, sodium pyruvate, and GlutaMAX (DMEM; ThermoFisher, 10569) supplemented with 10% Fetal Bovine Serum (FBS; ThermoFisher, 10438), and 100 U/mL Penicillin-Streptomycin (ThermoFisher, 15140).

Techniques: Flow Cytometry

Journal: bioRxiv

Article Title: Altered DNA repair pathway engagement by engineered CRISPR-Cas9 nucleases

doi: 10.1101/2022.03.10.483793

Figure Lengend Snippet: a , Precise editing and indel frequencies using HDR templates or PE3 prime editing at several loci. b , Precise editing and indel frequencies using MDR templates, PE2 prime editing, or PE3 prime editing at several loci in non-dividing (quiescent) primary human dermal fibroblasts. * indicates p < 0.05 for precise editing frequency compared to wild-type Cas9. Data were analyzed by deep next-generation sequencing and represent means of n = 3 independent replicates with standard errors.

Article Snippet: HEK293T human embryonic kidney, HeLa human cervical cancer, A549 human lung cancer, Panc1 human pancreatic cancer, and primary human dermal fibroblasts from neonates (GlobalStem) were maintained in Dulbecco’s Modified Eagle’s Medium with high glucose, sodium pyruvate, and GlutaMAX (DMEM; ThermoFisher, 10569) supplemented with 10% Fetal Bovine Serum (FBS; ThermoFisher, 10438), and 100 U/mL Penicillin-Streptomycin (ThermoFisher, 15140).

Techniques: Next-Generation Sequencing