Journal: iScience

Article Title: Reprogramming bone progenitor identity and potency through control of collagen density and oxygen tension

doi: 10.1016/j.isci.2022.104059

Figure Lengend Snippet: Development of a transitional culture to rescue AMSC skeletal stem cell phenotype (A) Schematic diagram illustrating the transitional culture. AMSCs were cultured in a 10% collagen type I gel for 7 days and subsequently introduced into a 0.2% collagen type I gel for a further 7 and 14 days. (B) Gene expression profile of progenitor cell markers were quantified using qPCR; human skeletal stem cell makers (CD164, PDPN, CD73), mesenchymal progenitor markers (NESTIN and PRX1), and a bone-cartilage skeletal progenitor marker (CD146); (C) Osteoblast and chondrocyte transcription factors RUNX2 and SOX9, respectively; and (D) Matrix stiffness associated markers (YAP, CTGF). (Data are presented as mean ±S.E.M, statistical analysis performed using one-way analysis of variance, uncorrected Fisher’s least significant difference; ∗∗∗p < 0.001; ∗∗p < 0.01; ∗p < 0.05; n = 3).

Article Snippet: Cells were stained with PDPN monoclonal (Thermo Fisher Scientific Cat# 12-9381-41, RRID: AB_1582263 , Loughborough, Leicestershire, UK), CD73 monoclonal (Thermo Fisher Scientific Cat# 11-0739-42, RRID: AB_10596508 ) and CD146 monoclonal antibodies (Thermo Fisher Scientific Cat# 11-1469-42, RRID: AB_2043805 ) for 1 h at room temperature.

Techniques: Cell Culture, Gene Expression, Marker

Journal: iScience

Article Title: Reprogramming bone progenitor identity and potency through control of collagen density and oxygen tension

doi: 10.1016/j.isci.2022.104059

Figure Lengend Snippet: Migration and differential protein expression of AMSCs from a high density collagen gel to a low density collagen gel (A) Transitional cultures stained with H&E. Cells were visible in the 0.2% collagen scaffold by day 14 (scale bar: 100 μm). (B) Immunofluorescent staining of CD73, PDPN, and CD146 of the transitional cultures. Red arrows indicate the 10% collagen type I gel; blue arrows indicate cells migrated into 0.2% collagen type I gel (scale bar: 50 μm, 20 μm).

Article Snippet: Cells were stained with PDPN monoclonal (Thermo Fisher Scientific Cat# 12-9381-41, RRID: AB_1582263 , Loughborough, Leicestershire, UK), CD73 monoclonal (Thermo Fisher Scientific Cat# 11-0739-42, RRID: AB_10596508 ) and CD146 monoclonal antibodies (Thermo Fisher Scientific Cat# 11-1469-42, RRID: AB_2043805 ) for 1 h at room temperature.

Techniques: Migration, Expressing, Staining

Journal: iScience

Article Title: Reprogramming bone progenitor identity and potency through control of collagen density and oxygen tension

doi: 10.1016/j.isci.2022.104059

Figure Lengend Snippet: Cell surface analysis of AMSCs within the transitional culture AMSCs were cultured in a 10% collagen type I gel for 7 days and embedded in a 0.2% collagen type I gel, making up the transitional culture for an additional 14 days. Flow cytometric analysis was conducted for PDPN, CD73, and CD146 (representative data presented).

Article Snippet: Cells were stained with PDPN monoclonal (Thermo Fisher Scientific Cat# 12-9381-41, RRID: AB_1582263 , Loughborough, Leicestershire, UK), CD73 monoclonal (Thermo Fisher Scientific Cat# 11-0739-42, RRID: AB_10596508 ) and CD146 monoclonal antibodies (Thermo Fisher Scientific Cat# 11-1469-42, RRID: AB_2043805 ) for 1 h at room temperature.

Techniques: Cell Culture

Journal: iScience

Article Title: Reprogramming bone progenitor identity and potency through control of collagen density and oxygen tension

doi: 10.1016/j.isci.2022.104059

Figure Lengend Snippet:

Article Snippet: Cells were stained with PDPN monoclonal (Thermo Fisher Scientific Cat# 12-9381-41, RRID: AB_1582263 , Loughborough, Leicestershire, UK), CD73 monoclonal (Thermo Fisher Scientific Cat# 11-0739-42, RRID: AB_10596508 ) and CD146 monoclonal antibodies (Thermo Fisher Scientific Cat# 11-1469-42, RRID: AB_2043805 ) for 1 h at room temperature.

Techniques: Recombinant, Plasmid Preparation, Derivative Assay, Reverse Transcription, SYBR Green Assay, Software