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Bovine fibrinogen

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Bovine fibrinogen is a purified protein derived from bovine blood plasma. It is a primary component in the blood clotting cascade and plays a crucial role in the formation of fibrin clots.

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Lab products found in correlation

Thrombin, by Merck Group (3 mentions) Bovine thrombin, by MP Biomedicals (2 mentions) Bovine plasma thrombin, by Abcam (1 mentions) Fibrinogen, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Ascorbic acid, by Merck Group (1 mentions) Aprotinin, by Merck Group (1 mentions) Penicillin streptomycin glutamine (psg), by Merck Group (1 mentions) Pbs 1x, by Thermo Fisher Scientific (1 mentions)

8 protocols using bovine fibrinogen

1

Evaluating 3D Cell Responses to Drugs

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A total of 200 trypsinized SW480 cells were mixed with 100 μl of 2.5 mg/ml bovine fibrinogen (MP Biomedicals) in DMEM plus 10% FBS and 1% penicillin–streptomycin–glutamine and 1 μl of thrombin (Sigma). The fibrin gels were seeded in 96‐well, flat‐bottom plates. After the gels solidified, 100 μl of DMEM media containing the desired drug treatment was layered on top (DCA was obtained from Sigma, XAV939 from Stemgent). Wells were imaged after 14 days of incubation. Size measurements were taken using Adobe Photoshop. Data were analyzed using Prism (GraphPad).
Lee M., Chen G.T., Puttock E., Wang K., Edwards R.A., Waterman M.L, & Lowengrub J. (2017). Mathematical modeling links Wnt signaling to emergent patterns of metabolism in colon cancer. Molecular Systems Biology, 13(2), 912.
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2

Fibrin Hydrogel for Cell Sheet Formation

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A provisional hydrogel was used to secure the cell sheet as it aggregated around the central post to form the ring structures. Hydrogels rapidly degrade in 2–4 weeks hence serving as a temporary support. Fibrin gel was identified as an ideal option as it is naturally found in the body. Fibrin hydrogels were formed using thrombin, fibrinogen, and hydrogel media28 (link). A 4:1 ratio of 20 mg/mL bovine fibrinogen (151122, MP Biomedicals LLC, OH) to 100 U/mL bovine plasma thrombin (7592, BioVision, Milpitas, CA) was prepared. Hydrogel media consisted of 88.8% DMEM, 0.1% TGF-β and ascorbic acid, 10% fetal bovine serum, and 1% antibiotic/antimycotic.
Patel B., Wonski B.T., Saliganan D.M., Rteil A., Kabbani L.S, & Lam M.T. (2021). Decellularized dermis extracellular matrix alloderm mechanically strengthens biological engineered tunica adventitia-based blood vessels. Scientific Reports, 11, 11384.
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3

Fibroblast-Keratinocyte Co-Culture for Skin Engineering

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Bovine fibrinogen (90% clottable, MP Biomedicals, Santa Ana, CA) was dissolved in 1.1% NaCl at 37°C for 4 hours and then filtered with a 0.45 μm nylon membrane filter. Fibroblasts were collected with the use of trypsin and a centrifuge, and re-suspended in media to a final concentration 2×106 cells per ml. 150 μl of this cell suspension was mixed with 1 ml of thrombin (3 IU - Sigma Aldrich, St. Louis, MO), and this cell/thrombin mix was added to fibrinogen at a ratio of 1:1. The mixture was quickly but gently distributed 1 ml/well into a 24-well plate and incubated at 37°C. After 20 minutes medium supplemented with ascorbic acid and aprotinin (Sigma, St. Louis, MO; final concentration 10 μg/ml) was added. The matrices were left to mature for 5–7 days while medium was changed every other day. Keratinocytes were plated on top at 2×106 per well and on the next day the culture was raised to the air-liquid interface on a metal grid and treatment with amlexanox was started; medium was changed every other day with fresh drug, ascorbic acid and aprotinin. Cultures were collected at one or two weeks of treatment and frozen with OCT in liquid nitrogen cooled isopentane. 8 μM sections were cut using a cryostat (AVANTIK QS11) and immunostained with the polyclonal type VII collagen antibody at a dilution of 1:800. Nuclei were counterstained with DAPI (Invitrogen, Carlsbad, CA).
Atanasova V.S., Jiang Q., Prisco M., Gruber C., Hofbauer J.P., Chen M., Has C., Bruckner-Tuderman L., McGrath J.A., Uitto J, & South A.P. (2017). Amlexanox enhances premature termination codon read-through in COL7A1 and expression of full length type VII collagen: potential therapy for recessive dystrophic epidermolysis bullosa. The Journal of investigative dermatology, 137(9), 1842-1849.
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4

Patterned Supported Lipid Bilayers

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A 1.5 μm thick sheet of polydimethylsiloxane polymer (PDMS) was produced by curing the polymer between two hydrophobized glass slides. A hole was cut into the center of the PDMS sheet, and a well was made by pressing it onto a clean glass coverslip. A PDMS stamp, which was cured over a seven-channel microfluidic pattern, was incubated in a 1 mg/mL bovine fibrinogen (MP Biomedicals, Solon, OH) solution and then pressed onto the glass coverslip in the well for 5 min before being removed. This left a patterned bovine fibrinogen imprint on the glass. Vesicles were then deposited in the well and incubated for 10 min before the excess was rinsed away with copious amounts of water. Continuous supported bilayers only formed in the regions lacking the bovine fibrinogen pattern. A second sheet of PDMS with an inlet and an outlet hole was then placed on top of the well, and the entire structure was held together by aluminum plates with two clamps to make a flow cell. Finally, fresh solution was flowed through the device as experiments were performed.
Robison A.D., Sun S., Poyton M.F., Johnson G.A., Pellois J.P., Jungwirth P., Vazdar M, & Cremer P.S. (2016). Polyarginine Interacts More Strongly and Cooperatively than Polylysine with Phospholipid Bilayers. The journal of physical chemistry. B, 120(35), 9287-9296.
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5

Fibrin Gel Culturing of Trypsinized Cells

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Briefly, 200 trypsinized cells were mixed with 100μL of 2.5 mg/mL bovine fibrinogen (MP Biomedicals) in DMEM plus 10% FBS and 1% Penicillin-Streptomycin-Glutamine and 1μL of thrombin (Sigma). The fibrin gels were seeded in 96 well, flat bottom plates. After the gels solidified, 100μL of DMEM media was layered on top. Wells were imaged regularly up to 14 days. Size measurements were taken using Adobe Photoshop. Data was analyzed using Prism (Graphpad, RRID:SCR_002798).
Chen G.T., Tifrea D.F., Murad R., Habowski A.N., Lyou Y., Duong M.R., Hosohama L., Mortazavi A., Edwards R.A, & Waterman M.L. (2022). Disruption of beta-catenin dependent Wnt signaling in colon cancer cells remodels the microenvironment to promote tumor invasion. Molecular cancer research : MCR, 20(3), 468-484.
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6

Fabrication of Fibrin Test Strips

Cited 1 time
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For fibrin test strip fabrication, 40 mg/mL bovine fibrinogen (MP Biomedicals) was combined with 10 U/mL bovine thrombin (MP Biomedicals) at a final concentration of 36 mg/mL and 1 U/mL, respectively, to catalyze the conversion of fibrinogen to fibrin67 (link). The fibrin mixture was casted into 3D printed plastic dog bone molds containing small Velcro pieces (VELCRO® Super-Grip Double-Head Hook) inserted at both ends to grip the gel during tensile testing. Molds were sprayed with Teflon (WD-40® SpecialistTM Dirt & Dust Resistant Dry Lube Spray) to facilitate mold release following casting. After 30 min at room temperature, the fibrin hydrogel solidified.
Shiwarski D.J., Tashman J.W., Tsamis A., Bliley J.M., Blundon M.A., Aranda-Michel E., Jallerat Q., Szymanski J.M., McCartney B.M, & Feinberg A.W. (2020). Fibronectin-based nanomechanical biosensors to map 3D surface strains in live cells and tissue. Nature Communications, 11, 5883.
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7

Fibrin-based Gel Scaffold for Cell Seeding

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Fibrin-based gels were prepared by mixing 300 µL of bovine fibrinogen (MP Biomedicals, Irvine, CA, USA) solution at 6 mg/mL in PBS 1X (Gibco) with 300 µL of high-glucose DMEM in the culture wells. Then, 300 µL of a 25 UI/mL thrombin solution (Merck, Rahway, NJ, USA) in pure H2O was added for polymerization. The fibrin-based gel was left to solidify for 45 min at 37 °C.
In configuration (i), the cells were directly seeded on the ceramic pellet at 20,000 cells/cm2 and incubated for 1 h. Afterwards, the pellets were transferred to another well, and the fibrin gel was layered onto the cell-seeded ceramic surface. After solidification, 1 mL DMEM was added in the well for cell cultivation. In configuration (ii), fibrin-based gels were directly poured on the pellets. After, solidification, the C166 cells were seeded on the gel at 20,000 cells/cm2 in 1 mL of complete culture medium.
The cells were cultured for 7 days, and the culture medium was renewed after 3 days. After 7 days, cells were fixed with 4% PFA for 10 min at RT.
Usseglio J., Dumur A., Pagès E., Renaudie É., Abélanet A., Brie J., Champion É, & Magnaudeix A. (2023). Microporous Hydroxyapatite-Based Ceramics Alter the Physiology of Endothelial Cells through Physical and Chemical Cues. Journal of Functional Biomaterials, 14(9), 460.
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8

Fabrication of Fibrin Test Strips

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For fibrin test strip fabrication, 40 mg/mL bovine fibrinogen (MP Biomedicals) was combined with 10 U/mL bovine thrombin (MP Biomedicals) at a final concentration of 36 mg/mL and 1 U/mL, respectively, to catalyze the conversion of fibrinogen to fibrin 52 (link) . The fibrin mixture was casted into 3D printed plastic dog bone molds containing small Velcro pieces (VELCRO® Super-Grip Double-Head Hook) inserted at both ends to grip the gel during tensile testing. Molds were sprayed with Teflon (WD-40® SpecialistTM Dirt & Dust Resistant Dry Lube Spray) to facilitate mold release following casting. After 30 minutes at room temperature, the fibrin hydrogel solidified.
Shiwarski D.J., Tashman J.W., Tsamis A., Bliley J.M., Blundon M.A., Aranda‐Michel E., Jallerat Q., Szymanski J.M., McCartney B.M., & Feinberg A.W. (2020). Fibronectin-Based Nanomechanical Biosensors to Map 3D Strains in Live Cells and Tissues.
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