Journal: Journal of Tissue Engineering

Article Title: Blood vessels bioengineered from induced pluripotent stem cell derived mesenchymal stem cells and porous silk fibroin coated functional scaffolds

doi: 10.1177/20417314251355723

Figure Lengend Snippet: hiPSC differentiate into cells with an MSC phenotype exhibiting common MSC markers. A schematic of the MSC differentiation protocol is shown (a) (Created with BioRender.com ). Fold gene expression increases in MSC marker-genes THY1 (CD90), NT5E (CD73) and ENG (CD105) was observed throughout differentiation of hiPSC-iMSCs (b). Histograms for common MSC positive markers are displayed (c) and increase of the percentage of cells positive for CD90, CD73 and CD105 after 36 days could also be noticed via flow cytometry (C-i to C-iii) after 36 days when iMSCs were derived. Homogenous iMSC populations comparable to hBM-MSCs positive for phenotypical MSCs markers CD44 and CD73 (green) as well as CD105 and CD90 (red) could be observed by day 34 of differentiation via Immunofluorescence staining (d). Scale bars shown at 100 μm. Data significance is presented as *** p ⩽ 0.001 and **** p ⩽ 0.0001 ( n = 3).

Article Snippet: Human bone marrow derived mesenchymal stem cells (hBM-MSC) (C-12974) harvested from normal human bone marrow from individual donors were cultured with Mesenchymal Stem Cell Growth Medium 2 (C-28009) purchased from PromoCell (Germany).

Techniques: Gene Expression, Marker, Flow Cytometry, Derivative Assay, Immunofluorescence, Staining

Journal: Journal of Tissue Engineering

Article Title: Blood vessels bioengineered from induced pluripotent stem cell derived mesenchymal stem cells and porous silk fibroin coated functional scaffolds

doi: 10.1177/20417314251355723

Figure Lengend Snippet: Characterisation of iMSC and hBM-MSCs via alizarin red staining. Both iMSCs (a-i) and hBM-MSCs (j-l) underwent osteogenic differentiation with noticeable mineralisation nodules being produced by the end of differentiation (day 28) as displayed by alizarin red staining. Images were collected on an Olympus IX83 inverted microscope and captured through MMI CellTools software at 10× magnification. Scale bars shown at 200 μm.

Article Snippet: Human bone marrow derived mesenchymal stem cells (hBM-MSC) (C-12974) harvested from normal human bone marrow from individual donors were cultured with Mesenchymal Stem Cell Growth Medium 2 (C-28009) purchased from PromoCell (Germany).

Techniques: Staining, Produced, Inverted Microscopy, Software

Journal: Journal of Tissue Engineering

Article Title: Blood vessels bioengineered from induced pluripotent stem cell derived mesenchymal stem cells and porous silk fibroin coated functional scaffolds

doi: 10.1177/20417314251355723

Figure Lengend Snippet: Proliferation and metabolic activity of hBM-MSC and iMSCs. A diagram of the modifications made to the pristine (1) scaffolds by immersing in acetone ⩾ 99.8% acetone to produce porous (2) fibres and further covered with a 1% w/v silk fibroin solution to coat scaffolds (3) (a) (Created with BioRender.com ). The modifications to the scaffolds were assessed to evaluate their biocompatibility compared to the control pristine elcetrospun scaffolds. The three different types of scaffolds were visualised under SEM imaging. Pristine scaffolds produced by electrospinning without any post-electrospinning modification (b). Cells count differences in iMSCs and hBM-MSCs seeded on pristine, porous and silk fibroin coated scaffolds after and reduction of AlamarBlue™ differences in iMSCs and hBM-MSCs seeded on pristine, porous and silk fibroin coated scaffolds after 10 days of culture (c) Density and morphological differences between the scaffold conditions were also observed via immunofluorescence staining with Phalloidin-iFluor 488 (a). Scale bars shown at 100 μm (d). Data significance is presented as * p < 0.05, ** p ⩽ 0.01, *** p ⩽ 0.001 ( n = 3).

Article Snippet: Human bone marrow derived mesenchymal stem cells (hBM-MSC) (C-12974) harvested from normal human bone marrow from individual donors were cultured with Mesenchymal Stem Cell Growth Medium 2 (C-28009) purchased from PromoCell (Germany).

Techniques: Activity Assay, Control, Imaging, Produced, Modification, Immunofluorescence, Staining

Journal: Journal of Tissue Engineering

Article Title: Blood vessels bioengineered from induced pluripotent stem cell derived mesenchymal stem cells and porous silk fibroin coated functional scaffolds

doi: 10.1177/20417314251355723

Figure Lengend Snippet: Vessel-like constructs derived from hBM-MSCs and iMSCs. Diagram of the fabrication process of tissue engineered blood vessels (a) (Created with BioRender.com ). The tube construct post-production can be seen next to the steel rod used to fabricate next and next to a penny for scale comparison (bi). Length dimensions (bii), wall thickness and inner diameter dimensions (biii) are also displayed. Immunofluorescence images of longitudinal cross-sections of tube constructs (b). Vessel mimics fabricated using both hBM-MSC-VSMCs and iMSC-VSMCs can be observed to be densely-populated with cells positive for α-SMA (green) and CNN1 (red). DAPI was counterstained to show nuclei. Scale bars shown at 100 μm. The representation of the scale is displayed in (c) Mechanical property differences in UTS (d), burst strength (e), young’s modulus (f) and strain (g) were also measured. Data significance is presented as * p < 0.05 ( n = 3).

Article Snippet: Human bone marrow derived mesenchymal stem cells (hBM-MSC) (C-12974) harvested from normal human bone marrow from individual donors were cultured with Mesenchymal Stem Cell Growth Medium 2 (C-28009) purchased from PromoCell (Germany).

Techniques: Construct, Derivative Assay, Comparison, Immunofluorescence