Journal: Journal of Functional Biomaterials

Article Title: Bovine Fibroblast-Derived Extracellular Matrix Promotes the Growth and Preserves the Stemness of Bovine Stromal Cells during In Vitro Expansion

doi: 10.3390/jfb14040218

Figure Lengend Snippet: The expansion of two batches of BUSC on BF-ECM or TCP for three passages. Batch 1 BUSCs were expanded from P3 to P6 on BF-ECM (ECM) or TCP for 24 days. The fold of amplification and doubling time were calculated for each passage (6 wells of BF-ECM and 6 wells of TCP) as described in Materials and Methods. The mean ± SD of three samples for each condition is shown. The statistical difference ( p -value) between ECM and TCP for each passage was determined by a student t -test. The total theoretical expansion (fold) from P3 to P6 was calculated by multiplying the folds of expansion of all 3 passages. Batch 2 BUSCs were expanded on BF-ECM or TCP similarly for 21 days.

Article Snippet: To evaluate the multipotency of isolated BUSCs, 2 × 10 4 /well of BUSCs were seeded to 24-well TCP plates (Cat #229124, CELLTREAT, Pepperell, MA, USA).

Techniques: Amplification

Journal: Journal of Functional Biomaterials

Article Title: Bovine Fibroblast-Derived Extracellular Matrix Promotes the Growth and Preserves the Stemness of Bovine Stromal Cells during In Vitro Expansion

doi: 10.3390/jfb14040218

Figure Lengend Snippet: BUSCs cultured on BF-ECM and TCP for multiple passages. BUSCs at different passages were seeded at 5 × 10 4 /well on BF-ECM (upper panel) or TCP (lower panel) 6-well plates. After being cultured for 7 days, phase contrast images were taken. Representative images were shown. Scale bar = 100 μm.

Article Snippet: To evaluate the multipotency of isolated BUSCs, 2 × 10 4 /well of BUSCs were seeded to 24-well TCP plates (Cat #229124, CELLTREAT, Pepperell, MA, USA).

Techniques: Cell Culture

Journal: Journal of Functional Biomaterials

Article Title: Bovine Fibroblast-Derived Extracellular Matrix Promotes the Growth and Preserves the Stemness of Bovine Stromal Cells during In Vitro Expansion

doi: 10.3390/jfb14040218

Figure Lengend Snippet: The growth of BUSCs at different passages on BF-ECM versus TCP. The cell viability of BUSCs at P3 cultured on BF-ECM (ECM) or TCP was monitored for 9 days. The % of viability at each time point was normalized to the viability on Day 1. % of growth = viability (Day N) /viability (Day 1) × 100% ( A ). The P3 cells on BF-ECM or TCP were collected and counted on Day 9 ( B ). The viability of cells at P5 was monitored for 11 days. Data shown are mean ± SD ( n = 4) ( C ). Statistical differences between ECM and TCP are indicated for each time point * p < 0.05, ** p < 0.01 *** p < 0.001.

Article Snippet: To evaluate the multipotency of isolated BUSCs, 2 × 10 4 /well of BUSCs were seeded to 24-well TCP plates (Cat #229124, CELLTREAT, Pepperell, MA, USA).

Techniques: Cell Culture

Journal: Journal of Functional Biomaterials

Article Title: Bovine Fibroblast-Derived Extracellular Matrix Promotes the Growth and Preserves the Stemness of Bovine Stromal Cells during In Vitro Expansion

doi: 10.3390/jfb14040218

Figure Lengend Snippet: The growth of BUSCs (P3) on BF-ECM (ECM) versus TCP in the presence of different percentages of bovine serum. ( A ) The cell viability was measured using the alamarBlue assay. % of growth = viability (Day N) /viability (Day 1) × 100%. Data shown are mean ± SD ( n = 3). ( B ) The statistical differences ( p values) between different culturing conditions at each time point were determined using one-way ANOVA with Tukey’s multiple comparisons tests (GraphPad Prism).

Article Snippet: To evaluate the multipotency of isolated BUSCs, 2 × 10 4 /well of BUSCs were seeded to 24-well TCP plates (Cat #229124, CELLTREAT, Pepperell, MA, USA).

Techniques: Alamar Blue Assay

Journal: Journal of Functional Biomaterials

Article Title: Bovine Fibroblast-Derived Extracellular Matrix Promotes the Growth and Preserves the Stemness of Bovine Stromal Cells during In Vitro Expansion

doi: 10.3390/jfb14040218

Figure Lengend Snippet: Adipogenic differentiation of BUSCs expanded on TCP or BF-ECM with an optimized adipogenic medium. BUSCs were expanded on TCP or BF-ECM (ECM) till passage 8. Cells (at P9) were seeded on TCP, induced with an optimized adipogenic medium for 7 days, and stained with Oil Red O ( A ) Scale bar = 50 μm. ( B ) Oil red O dye was extracted from stained cells and quantified and normalized to the number of cells (μg of DNA). Data shown are mean ± SD ( n = 3) ** p < 0.01.

Article Snippet: To evaluate the multipotency of isolated BUSCs, 2 × 10 4 /well of BUSCs were seeded to 24-well TCP plates (Cat #229124, CELLTREAT, Pepperell, MA, USA).

Techniques: Staining

Journal: ACS Applied Bio Materials

Article Title: Differential Effects of Confinement on the Dynamics of Normal and Tumor-Derived Pancreatic Ductal Organoids

doi: 10.1021/acsabm.4c01301

Figure Lengend Snippet: Microgel composites increase confinement. (A) Illustration of microgel processing from bulk polyacrylamide hydrogel to 100 μm diameter microgels mixed with Matrigel for organoid culture. (B) Parallel plate rheometer geometry used to characterize the rheological properties of microgel composites. (C) Average storage modulus ( G ′) for Matrigel (less confined, n = 3) and 75 vol % microgel and 25 vol % Matrigel composites (more confined, n = 2). (D) Average yield stress of less confined ( n = 3) and more confined ( n = 2) microenvironment. (E) Confocal maximum intensity projection of confined microenvironment with FITC-Matrigel (green) and microgels labeled with orange polystyrene nanoparticles (diameter = 500 nm, particle concentration = 0.01 wt %, Ex. 542 nm/Em. 612 nm). (F) 3D heat map of channel width between microgel particles showing average channel width ≈ 8 μm. (G) Confocal maximum intensity projection of an organoid midplane. Organoid stained with CellTracker CMFDA (cytosol) and DNA (Hoechst 33342) with a lumen in the center. Microgels embedded with orange fluorescent nanoparticles (diameter = 500 nm, particle concentration = 0.01 wt %) and outlined with white dashes. Statistical significance was determined from an unpaired t -test. * p ≤ 0.05. Error: SD. Scale bars = 100 μm.

Article Snippet: Pancreas ductal organoids were maintained in Matrigel-dome culture up to passage 40 in GenClone tissue culture treated, 24-well, flat-bottom polystyrene plates (Genesee Scientific, cat.#25–107).

Techniques: Labeling, Concentration Assay, Staining

Journal: STAR Protocols

Article Title: Protocol to Fabricate Engineered Illumination Devices for Optogenetic Control of Cellular Signaling Dynamics

doi: 10.1016/j.xpro.2020.100141

Figure Lengend Snippet:

Article Snippet: VisiPlate-24 Black, Black 24-well Microplate with Clear Bottom , Perkin Elmer , Cat#1450-605.

Techniques: Software, Blocking Assay, Cell Culture, Imaging