human bone marrow derived mscs c 12974  (PromoCell)

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

Journal: Scientific Reports

Article Title: Bioengineering of vascularized porcine flaps using perfusion-recellularization

doi: 10.1038/s41598-024-58095-7

Figure Lengend Snippet: Recellularization of acellular omentum and tensor fascia lata flaps. Forty million HUVEC and forty million MSCs were manually seeded with syringe via the arterial inlet. Following 4 h of static culture, scaffolds were perfused with EGM2 growth media for a total culture period of 6 days. Evidence of cell attachment as noted in the recellularized omentum ( A ) and the tensor fascia lata ( B ) at the interface of the vascular lumen (asterisks) determined using H&E and DAPI visualization. H&E stains of small and relatively larger caliber vessels are depicted in top and middle row, respectively. Images are representative of n = 3 of each condition: native, decellularized and recellularized tissues. Scale Bar: 200 µm.

Article Snippet: Commercially obtained human bone-marrow derived MSCs (Promocell, Germany) were cultured in MSCGM (Promocell) containing proprietary media supplement and 5% FBS.

Techniques: Cell Attachment Assay

Journal: Scientific Reports

Article Title: Bioengineering of vascularized porcine flaps using perfusion-recellularization

doi: 10.1038/s41598-024-58095-7

Figure Lengend Snippet: Recellularization with HUVEC and MSC co-culture regenerates endothelial phenotype in decellularized omentum and TFL flaps. Forty million HUVEC and forty million MSCs were manually seeded with syringe via the arterial inlet and cultured in bioreactor for a total of 6 days. CD31 positivity was demonstrated using IHC at the interface of the vascular lumen (asterisks) in the recellularized condition. No CD31was seen in decellularized control. Representative Images from n = 3 samples. Scale Bar: 200 µm.

Article Snippet: Commercially obtained human bone-marrow derived MSCs (Promocell, Germany) were cultured in MSCGM (Promocell) containing proprietary media supplement and 5% FBS.

Techniques: Co-Culture Assay, Cell Culture, Control

Journal: Regenerative therapy

Article Title: Generation of functional liver sinusoidal endothelial-like cells from human bone marrow-derived mesenchymal stem cells.

doi: 10.1016/j.reth.2023.07.006

Figure Lengend Snippet: Fig. 1. Screening of cytokines and small molecules for LSEC differentiation. (A) The schematic representation of BM-MSC differentiation. (B) qPCR analysis of the expression levels of LYVE1 and CD36 in BM-MSC-derived cells. On the y-axis, the expression levels are shown as a relative value to those of BM-MSCs. All data are presented as mean ± SD (n ¼ 3). Significant differences were evaluated using a one-way ANOVA followed by Dunnett's post-hoc test (*p < 0.05, ***p < 0.001 compared with BFTc [BMP4, FGF8b, TGF-b inhibitor, and cAMP]). Abbreviations: B; BMP4, F; FGF8b, T; TGF-b signal inhibitor, c; cAMP.

Article Snippet: Primary human bone marrow-derived MSCs (C-12974; PromoCell) were cultured in Mesenchymal Stem Cell Growth Medium 2 (C-28009; PromoCell).

Techniques: Expressing, Derivative Assay

Journal: Regenerative therapy

Article Title: Generation of functional liver sinusoidal endothelial-like cells from human bone marrow-derived mesenchymal stem cells.

doi: 10.1016/j.reth.2023.07.006

Figure Lengend Snippet: Fig. 2. Analysis of the expression of LSEC-related markers. (A) The schematic representation of BM-MSC differentiation to LSEC-like cells. (B) Brightfield images of the BM-MSCs and BM-MSC-derived cells. Scale bar ¼ 500 mm. (C) qPCR analysis of gene expression of LYVE1, CD36, CD32b, F8, PLVAP, CD31, and VEGFR2 in the BM-MSCs and BM-MSC-derived cells. On the y-axis, the expression levels are shown as a relative value to those of BM-MSCs. Data are presented as mean ± SD (n ¼ 6, sum of two independent experiments, n ¼ 3 for each experiment). (D) Immunocytochemistry analysis of LYVE1 (green) and CD36 (red) in the BM-MSCs and BM-MSC-derived cells. The nuclei were counterstained with DAPI (blue). Scale bar ¼ 50 mm. (E) The expression level of CD32 in the BM-MSCs and BM-MSC-derived cells was measured using flow cytometry analysis. The values of mean fluorescence intensity (MFI) of CD32 expression were normalized by the MFI of isotype control. Data are presented as mean ± SD (n ¼ 3). Significant differences were evaluated using an unpaired two-tailed Student's t-test (***p < 0.001). (F) The percentage of LYVE1- or CD36-positive cells in the BM-MSC-derived cells was measured using flow cytometry analysis.

Article Snippet: Primary human bone marrow-derived MSCs (C-12974; PromoCell) were cultured in Mesenchymal Stem Cell Growth Medium 2 (C-28009; PromoCell).

Techniques: Expressing, Derivative Assay, Gene Expression, Immunocytochemistry, Cytometry, Control, Two Tailed Test

Journal: Regenerative therapy

Article Title: Generation of functional liver sinusoidal endothelial-like cells from human bone marrow-derived mesenchymal stem cells.

doi: 10.1016/j.reth.2023.07.006

Figure Lengend Snippet: Fig. 3. Evaluation of endothelial cell-related functions. (A) Tube-formation assay of the BM-MSCs, BM-MSC-derived cells, and HUVECs. Scale bar ¼ 200 mm. (B) acLDL-uptake assay. BM-MSCs, BM-MSC-derived cells, and HUVECs were cultured in a medium containing Alexa Fluor 488-conjugated acLDL to measure their ability to take up acLDL. Scale bar ¼ 20 mm. Abbreviation: LDL, low-density lipoprotein.

Article Snippet: Primary human bone marrow-derived MSCs (C-12974; PromoCell) were cultured in Mesenchymal Stem Cell Growth Medium 2 (C-28009; PromoCell).

Techniques: Tube Formation Assay, Derivative Assay, Cell Culture

Journal: Regenerative therapy

Article Title: Generation of functional liver sinusoidal endothelial-like cells from human bone marrow-derived mesenchymal stem cells.

doi: 10.1016/j.reth.2023.07.006

Figure Lengend Snippet: Fig. 4. Evaluation of LSEC-related functions. (A) HA-uptake assay. BM-MSCs, BM-MSC-derived cells, and HUVECs were cultured in a medium containing fluoresceinamine- conjugated HA to measure their ability to take up HA. (B) IgG-uptake assay. BM-MSCs, BM-MSC-derived cells, and HUVECs were cultured in a medium containing Alexa Fluor 488-conjugated IgG to measure their ability to endocytose IgG. Scale bar ¼ 20 mm. Abbreviation: HA, hyaluronic acid.

Article Snippet: Primary human bone marrow-derived MSCs (C-12974; PromoCell) were cultured in Mesenchymal Stem Cell Growth Medium 2 (C-28009; PromoCell).

Techniques: Derivative Assay, Cell Culture

Journal: Regenerative therapy

Article Title: Generation of functional liver sinusoidal endothelial-like cells from human bone marrow-derived mesenchymal stem cells.

doi: 10.1016/j.reth.2023.07.006

Figure Lengend Snippet: Fig. 5. Evaluation of the differentiation capacity of primary human BM-MSCs into LSEC-like cells (A) The schematic representation of the differentiation of primary human BM- MSCs into LSEC-like cells. (B) qPCR analysis of gene expression of LYVE1, CD36, CD32b, and F8 in primary human BM-MSCs and primary human BM-MSC-derived cells. On the y-axis, the expression levels are shown as a relative value to those of primary human BM-MSCs. Data are presented as mean ± SD (n 8, sum of two independent experiments, n 3 for each experiment.). Significant differences were evaluated using an unpaired two-tailed Student's t-test (*p < 0.05, ***p < 0.001). (C) The percentage of LYVE1- or CD36- positive cells in the primary human BM-MSC-derived cells was measured using flow cytometry analysis. Data are presented as mean ± SD (n ¼ 3). (D) acLDL-uptake assay. BM-MSCs, BM-MSC-derived cells, and HUVECs were cultured in a medium containing Alexa Fluor 488-conjugated acLDL to measure their ability to take up acLDL. Scale bar ¼ 20 mm. (E) HA- uptake assay. Primary human BM-MSCs and primary human BM-MSC-derived cells were cultured in a medium containing fluoresceinamine-conjugated HA to measure their ability to take up HA. Scale bar ¼ 20 mm. Abbreviation: LDL, low-density lipoprotein; HA, hyaluronic acid.

Article Snippet: Primary human bone marrow-derived MSCs (C-12974; PromoCell) were cultured in Mesenchymal Stem Cell Growth Medium 2 (C-28009; PromoCell).

Techniques: Gene Expression, Derivative Assay, Expressing, Two Tailed Test, Cytometry, Cell Culture