human adipose derived mesenchymal stem cells asc  (ATCC)

Journal: Journal of Biological Engineering

Article Title: Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow

doi: 10.1186/s13036-025-00491-2

Figure Lengend Snippet: Human ASC osteogenesis is supported by culture on SBG-PLGA composites and rhBMP-2 treatment. mRNA levels of the early and late osteoblastic markers, BMP-2 and Noggin ( NOG ) in human ASCs cultured on SBG-PLGA composites in ( A ), ( C ) standard osteogenic medium or ( B ), ( D ) standard osteogenic medium supplemented with 100 ng/ml rhBMP-2. Results are presented as relative mRNA expression levels vs. mRNA levels for ASCs cultured on a plain PLGA control (black line at 1). ( E ) Nitric oxide (NO) concentration in culture media after 24-h culture of ASC cells on SBG-PLGA composites in standard osteogenic medium. Averages ± SD are indicated. One-way or two-way ANOVA tests, * p < 0.05, ** p < 0.001, *** p < 0.0001 relative to the PLGA control group

Article Snippet: ASC52telo cells (ASC; ATCC, SCRC-4000) and normal human ASCs (ATCC, PCS-500-011) were expanded in the dedicated medium (ATCC, Mesenchymal Stem Cell Basal Medium with Mesenchymal Stem Cell Growth Kit and G418).

Techniques: Cell Culture, Expressing, Control, Concentration Assay

Journal: Journal of Biological Engineering

Article Title: Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow

doi: 10.1186/s13036-025-00491-2

Figure Lengend Snippet: Cumulative osteogenic effect of Phenamil and PD98059 treatment in rhBMP-2 stimulated human ASCs cultured on SBG-PLGA composites. mRNA levels of osteoblastic markers in ( A ) 7-day and ( B ) 21-day ASC cultures on SBG-PLGA composites. ASCs were cultured in osteogenic medium supplemented with 100 ng/ml rhBMP-2 or 100 ng/ml rhBMP-2, 50 µM PD98059 and 20 µM Phenamil. Results are presented as relative mRNA expression compared to mRNA levels in control cells cultured on PLGA with rhBMP-2 only (marked as black line at 1). ( C ) mRNA levels of selected osteoblastic markers in 3-day osteogenic ASC cultures treated with different doses of rhBMP-2 (25–250 ng/ml), Phenamil (5–50 µM) or PD98059 (1-125 µM); under fluid shear stress. Results are presented as the expression relative to osteogenic cultures treated only with ascorbic acid, dexamethasone and β-glycerophosphate. ( D ) Graphical hypothesis of BMP-2, PD98059 and Phenamil cross-talk in intracellular signaling. Average values ± SD are indicated. Two-way ANOVA test, * p < 0.05, ** p < 0.001, *** p < 0.0001 relative to the PLGA control group or between marked groups. BMP-2 – bone morphogenetic protein 2, OC – osteocalcin, ON – osteonectin, FOS – AP-1 transcription factor subunit (c-fos), OPG – osteoprotegerin

Article Snippet: ASC52telo cells (ASC; ATCC, SCRC-4000) and normal human ASCs (ATCC, PCS-500-011) were expanded in the dedicated medium (ATCC, Mesenchymal Stem Cell Basal Medium with Mesenchymal Stem Cell Growth Kit and G418).

Techniques: Cell Culture, Expressing, Control, Shear

Journal: Journal of Biological Engineering

Article Title: Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow

doi: 10.1186/s13036-025-00491-2

Figure Lengend Snippet: Fluid shear stress strengthens the osteogenic effects of rhBMP-2, PD98059 and Phenamil in ASCs cultured on SBG-PLGA composites. mRNA levels of osteoblastic markers after 7-day ASC culture on SBG-PLGA composites in ( A ) standard osteogenic medium under either static conditions or with fluid shear stress; and ( C ) osteogenic medium supplemented with 100 ng/ml rhBMP-2, 50 µM PD98059 and 20 µM Phenamil under either static conditions or with fluid shear stress. Results are presented as relative mRNA expression levels compared to mRNA levels in a control, static culture on PLGA (marked as a black line at 1). ( B ) The method of fluid shear stress application in ASC cultures using a standard laboratory see-saw rocker (7° tilt angle, 6 RPM frequency). ( D ) F-actin distribution in ASCs (Phalloidin-Atto488, magenta colored) at day 3 of culture in osteogenic medium supplemented with rhBMP-2, PD98059 and Phenamil after continuous static or dynamic culture conditions applied for 3 days. Scale bar represents 100 μm. ( E ) Western blot (WB) analysis of p-ERK1/2 and p-SMAD1/5/8 in ASCs after 1-h treatment with rhBMP-2 or rhBMP-2, PD98059 and Phenamil in static or dynamic conditions (upper panel) along with densitometric quantifications of WB results normalized to GAPDH levels. Averages ± SD are indicated. Two-way ANOVA test, * p < 0.05, ** p < 0.001, *** p < 0.0001 relative to the static PLGA control or between marked groups

Article Snippet: ASC52telo cells (ASC; ATCC, SCRC-4000) and normal human ASCs (ATCC, PCS-500-011) were expanded in the dedicated medium (ATCC, Mesenchymal Stem Cell Basal Medium with Mesenchymal Stem Cell Growth Kit and G418).

Techniques: Shear, Cell Culture, Expressing, Control, Western Blot

Journal: Journal of Biological Engineering

Article Title: Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow

doi: 10.1186/s13036-025-00491-2

Figure Lengend Snippet: Zinc (ZnO) or strontium (SrO) modified SBG-PLGA composites combined with fluid shear stress and BMP-based chemical stimulation, further increase osteogenic markers expression in early ASC cultures. mRNA levels of osteoblastic markers in ( A ) 3-day and ( B ) 6-day osteogenic ASC cultures on PLGA-based composites containing either unmodified or SrO- or ZnO-modified SBGs. Cells were treated with a combination of rhBMP-2, PD98059 and Phenamil at the indicated culture times in either static cultures or under fluid shear stress. Upper panels show the schemes of the ASC treatments. Results are presented as relative mRNA expression levels vs. mRNA levels in a control, static culture on PLGA (marked as a black line at 1). ( C ) Western blot (WB) analysis of phospho-β-catenin(Ser552), COX-2 and phospho-CREB(Ser133) levels in ASCs after 1-h treatment with rhBMP-2 or rhBMP-2, PD98059 and Phenamil in static or dynamic conditions (left panel) along with densitometric quantifications of WB results normalized to GAPDH levels (right panel). ( D ) Hypothesized signaling pathways involved in osteogenic response to treatment strategy. Averages ± SD are indicated. Two-way ANOVA test, * p < 0.05, ** p < 0.001, *** p < 0.0001 relative to the respective static PLGA control group or between marked groups

Article Snippet: ASC52telo cells (ASC; ATCC, SCRC-4000) and normal human ASCs (ATCC, PCS-500-011) were expanded in the dedicated medium (ATCC, Mesenchymal Stem Cell Basal Medium with Mesenchymal Stem Cell Growth Kit and G418).

Techniques: Modification, Shear, Expressing, Control, Western Blot, Protein-Protein interactions

Journal: Scientific reports

Article Title: Noggin promotes osteogenesis in human adipose-derived mesenchymal stem cells via FGFR2/Src/Akt and ERK signaling pathway.

doi: 10.1038/s41598-024-56858-w

Figure Lengend Snippet: Figure 1. Noggin protein increases alkaline phosphatase (ALP) activity and early osteogenic genes expression in adult human mesenchymal stem cells from different tissues. Alkaline phosphatase (ALP) activity after 7-d culture of (a) normal human bone marrow stromal cells (BMSCs), (b) normal human dental pulp stem cells (DPSCs) and (c) human immortalized adipose-derived stem cell line (ASC52telo). Cells were treated with either 100 ng/ml recombinant human Noggin (NOG) or 100 ng/ml recombinant human bone morphogenetic protein 2 (BMP-2), or both (BMP-2 + NOG), in osteogenic medium containing ascorbic acid (Asc) and dexamethasone (Dex). (d) ALP activity after 7-d culture of ASC52telo cells in osteogenic medium with different Noggin doses (100–400 ng/ml). (a–d) Control represents cells maintained in standard growth medium. Relative mRNA levels (qPCR) of selected osteoblastic markers in (e) normal human ASCs and (f) normal human BMSCs continuously treated with Noggin (NOG) or BMP-2 for 7 days in osteogenic medium. Relative quantification to control cells cultured in osteogenic medium. (a–f) Average values ± SD are plotted. One-way ANOVA tests, *p < 0.05, **p < 0.001, ***p < 0.0001, ns—not significant, relative to respective control or between marked groups.

Article Snippet: ASC52telo cells (ASC; ATCC, SCRC-4000) and normal human ASC (ATCC, PCS-500-011) were expanded in the dedicated medium (ATCC, Mesenchymal Stem Cell Basal Medium with Mesenchymal Stem Cell Growth Kit).

Techniques: Activity Assay, Expressing, Derivative Assay, Recombinant, Control, Quantitative Proteomics, Cell Culture

Journal: Scientific reports

Article Title: Noggin promotes osteogenesis in human adipose-derived mesenchymal stem cells via FGFR2/Src/Akt and ERK signaling pathway.

doi: 10.1038/s41598-024-56858-w

Figure Lengend Snippet: Figure 7. The suggested new signaling pathways induced by Noggin in human ASC osteogenic cultures. We have demonstrated that Noggin can activate FGFR2 receptors and Src kinase associated with the receptor complex. This results in ERK1/2 phosphorylation and, independently of PI3k, Akt kinase phosphorylation. It is known that dexamethasone, a component of osteogenic medium, stimulates RUNX2 and TAZ expressions. We have shown Noggin activation of Akt that leads to blocking the ability of GSK3 to degrade TAZ and suppress RUNX2 activity, thereby stabilizing TAZ and enhancing formation of RUNX2-TAZ complexes. Whereas RUNX2-TAZ complexes can be phosphorylated by Noggin-activated ERK1/2. Such activated RUNX2-TAZ complexes are required for the transcription of osteogenic genes. Besides, we have shown Noggin-related inhibition of SMAD1/5/8 activity, which may be a result of increased SMAD 7 expression due to Akt activity. Figure was created in Affinity Designer software (1.10.6).

Article Snippet: ASC52telo cells (ASC; ATCC, SCRC-4000) and normal human ASC (ATCC, PCS-500-011) were expanded in the dedicated medium (ATCC, Mesenchymal Stem Cell Basal Medium with Mesenchymal Stem Cell Growth Kit).

Techniques: Protein-Protein interactions, Phospho-proteomics, Activation Assay, Blocking Assay, Activity Assay, Inhibition, Expressing, Software