Journal: Scientific Reports

Article Title: Characterization of TGFβ1-induced tendon-like structure in the scaffold-free three-dimensional tendon cell culture system

doi: 10.1038/s41598-024-60221-4

Figure Lengend Snippet: Development of 3D tendon cell culture system. ( A ) Three major steps of 3D tendon culture: cell extraction, cell growth in monolayer, and 3D culture. ( B ) The timeline of monolayer tendon cell culture after cell extraction. ( C ) The base of the 3D tendon cell culture consists of a growth channel area molded into 2% agarose using 3D-printed-mold and cylindrical anchors wrapped by hydrophilized PCL. Primary cells from the mouse tail tendon were seeded (2.5 × 10 6 cells per construct) into the fibronectin-coated growth area to generate a 3D tendon construct. The dimension of growth area is 10 mm (length) × 1 mm (width) × 4 mm (depth). ( D ) The timeline of 3D tendon culture following monolayer culture (D, days after seeding of cells for 3D tendon culture; T, days after TGFβ1 treatment).

Article Snippet: The growth channels were then coated with Human plasma-derived fibronectin (0.375 mg/ml in 1X PBS, Corning) and dried for 20 min.

Techniques: Cell Culture, Extraction, Construct

Journal: ACS Omega

Article Title: Engineering a Hybrid Ti6Al4V-Based System for Responsive and Consistent Osteogenesis

doi: 10.1021/acsomega.3c07232

Figure Lengend Snippet: Fibronectin coating reduces variability in mineralization and when combined with hydrothermal treatment, it upregulates the expression of the gene associated with osteogenic differentiation. (a) Variability (standard deviation) in mineralization at different time points of bone marrow-derived human mesenchymal stem cells (MSCs) seeded on Ti6Al4V subjected to different treatments. (b) Relative gene expression levels of hallmarks of osteogenic differentiation in MSCs seeded on Ti6Al4V subjected to different treatments. Gene expression data are presented as fold changes at day 21 relative to the expression level on day 0.

Article Snippet: MSCs were seeded on Ti6Al4V groups with no treatment (Ti–O) and with hydrothermal treatment (Ti–H) and on both groups coated with human plasma-derived fibronectin (05-752-1, PRIME-XV human fibronectin; FUJIFILM Irvine Scientific, The Netherlands), corresponding to Ti–OC and Ti–HC, respectively.

Techniques: Expressing, Standard Deviation, Derivative Assay

Journal: bioRxiv

Article Title: Biocompatibility characterisation of CMOS-based Lab-on-Chip electrochemical sensors for in vitro cancer cell culture applications

doi: 10.1101/2023.11.23.568427

Figure Lengend Snippet: (a) MDA-MB-231 attached to ISFET arrays with deposited (i) Collagen Type I (50 µ g/ml), (ii) Fibronectin (1 µ g/ml) and (iii) PLOL (8 µ g/ml) deposited on ISFET surface prior to cell culture (from left to right). ISFETs are fixed and stained for cytoskeletal marker, α -Tubulin (red), proliferation marker, Ki67 (green), Hoescht nuclear dye (blue) in the upper panels and YAP/TAZ (yellow) in the lower panels. All images obtained on the Opera Phenix (Perkin Elmer). (b) Number of cells attached per well, after 48 hours, on all treated ISFET chips or untreated control chips versus standard culture plastic vessels. (c) Percentage of total population of cells attached on the respective ISFET chips whose shape were classified as elongated, star or round. (d) Kernel density estimation and corresponding rug plot for proliferation marker, Ki67, expression normalised to Hoescht dye intensity in cells cultured on control chips and treated chips. (e) Normalized mean Ki67/Hoescht intensity in treated chips or untreated chips versus standard culture plastic. (f) Kernel density estimation and corresponding rug plot for YAP/TAZ nuclear: nuclear ring ratio intensity on control chips or treated chips. (g) Normalized mean YAP/TAZ N:RR in treated chips or untreated chips versus standard culture plastic. N = 3,743 single cells for all KDE plots. All images obtained using the Opera Phenix (Perkin Elmer) and segmented with Columbus (Perkin Elmer). Kolmogorov-Smirnov statistics (Ks) performed using the scipy.stats module in Python.

Article Snippet: Fibronectin derived from human plasma (Sigma-Aldrich, Ref F0895) was diluted from a 0.1% solution to 1 µ g/ml with PBS.

Techniques: Cell Culture, Staining, Marker, Expressing

Journal: bioRxiv

Article Title: Biocompatibility characterisation of CMOS-based Lab-on-Chip electrochemical sensors for in vitro cancer cell culture applications

doi: 10.1101/2023.11.23.568427

Figure Lengend Snippet: (a) Cancerous cell line, U2OS CDK1 AS (osteosarcoma) was cultured on a standard tissue culture mattek dish (WT) or an ISFET array (Chip) both coated with fibronectin. In each panel: Right image shows paxillin stains of focal adhesions. Left image shows detection threshold segmentation of focal adhesions using the Focal Adhesion Analysis Server (FAAS). (b) Histogram of orientation of focal adhesions detection in WT versus chip conditions. (c) Mean area of single focal adhesions in WT versus on chip, as measured in square pixels. Images obtained on Total Internal Fluorescence (TIRF) microscope and quantiatively analysed using the Focal Adhesion Analysis Server. Graphs and statistics performed with GraphPad Prism version 9.5.0.

Article Snippet: Fibronectin derived from human plasma (Sigma-Aldrich, Ref F0895) was diluted from a 0.1% solution to 1 µ g/ml with PBS.

Techniques: Cell Culture, Fluorescence, Microscopy

Journal: bioRxiv

Article Title: Biocompatibility characterisation of CMOS-based Lab-on-Chip electrochemical sensors for in vitro cancer cell culture applications

doi: 10.1101/2023.11.23.568427

Figure Lengend Snippet: (a) MCF10A cells cultured on the Si 3 N 4 interface of ISFET chips and fluorescently stained for (i) cytoskeletal marker α -Tubulin (red), proliferation marker Ki67 (green), Hoescht nuclear dye (blue) (ii) YAP/TAZ (yellow). (b) Kernel density estimation and corresponding rug plot of cell ratio width to length for MCF10A cells cultured on 96-well standard plastic cell culture plates as a control or ISFET chips. (c) Cumulative distribution function of Ki67 intensity, normalized to nuclear Hoescht dye intensity in cells cultured on plastic or ISFET chips. Panel of kernel density estimation plot showing single cell population density. (d) Cumulative distribution function of YAP/TAZ nuclear to nuclear ring ratio intensity in cells cultured on plastic or ISFET chips. Panel of kernel density estimation plot showing single cell population density. (e) Number of cells attached per well for ISFET chips with extracellular matrix deposition of Collagen Type I (50 µ g/ml), Fibronectin (10ng/ml) and PLOL (8 µ g/ml) or control chips with no additional deposition. (f) Kernel density estimation and corresponding rug plot for proliferation marker, Ki67, intensity normalised to nuclear Hoescht dye intensity in MCF10A cells cultured on control ISFET chips versus ISFET chip configurations. (g) Kernel density estimation and corresponding rug plot for YAP/TAZ nucleus to nuclear ring ratio intensity in MCF10A cells cultured on control, ISFET chips versus ISFET chip configurations. N = 4,282 single cells for all plastic and control ISFET chip experiments. N = 2,119 single cells for all chip configurations. All images obtained using the Opera Phenix (Perkin Elmer) and segmented with Columbus (Perkin Elmer). Kolmogorov-Smirnov statistics (Ks) performed using the scipy.stats module in Python.

Article Snippet: Fibronectin derived from human plasma (Sigma-Aldrich, Ref F0895) was diluted from a 0.1% solution to 1 µ g/ml with PBS.

Techniques: Cell Culture, Staining, Marker