Bgjb medium

Manufactured by Thermo Fisher Scientific

Sourced in United States

BGJb medium is a cell culture medium designed for the in vitro cultivation of cells. It provides a balanced salt solution and essential nutrients required for the growth and maintenance of cell lines.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions) L ascorbic acid, by Merck Group (5 mentions) Penicillin, by Thermo Fisher Scientific (5 mentions) Streptomycin, by Thermo Fisher Scientific (5 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) Bovine serum albumin (bsa), by Merck Group (3 mentions) Low melting agarose, by Fujifilm (2 mentions) Ascorbic acid, by Thermo Fisher Scientific (2 mentions) A2356, by Merck Group (2 mentions) β glycerol phosphate, by Merck Group (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Pronase, by Merck Group (2 mentions) Dnase 1, by New England Biolabs (2 mentions) Facsaria 2, by BD (2 mentions) Cell strainer, by BD (2 mentions) Picmorg50, by Merck Group (2 mentions) Ratlaps ctx 1 eia kit, by Immunodiagnostic Systems (2 mentions) Ly294002, by MedChemExpress (1 mentions) 35 mm tissue culture dishes, by BD (1 mentions) Pan ras in 1, by MedChemExpress (1 mentions)

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BGJb culture medium (3 citations)

45 protocols using bgjb medium

Embryonic Craniofacial Explant Culture

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Timed-pregnant mice were euthanized at E13.5 and decapitated in PBS. The mandible and tongue were removed from the embryos, and each explant, including the upper half of the head, was placed in a glass tube containing BGJb medium (Gibco, 12591) supplemented with 50% fetal bovine serum, 0.1% ascorbic acid, and antibiotics. The tubes were placed in a rotary apparatus rotating at 50 rpm in an incubator at 37°C and 5% CO₂. After 3 days in culture with/without 30 μM atRA, the explants were fixed in 4% PFA and processed.

Yoshioka H., Mikami Y., Ramakrishnan S.S., Suzuki A, & Iwata J. (2021). MicroRNA-124-3p Plays a Crucial Role in Cleft Palate Induced by Retinoic Acid. Frontiers in Cell and Developmental Biology, 9, 621045.

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Dexamethasone-induced Osteoblast Calcification

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Cells were cultured for 7 days in BGJb medium (Fitton-Jackson Modification, Gibco, 12591038) (plus 10% FBS, 50 mg/mL ascorbic acid, 20 mM β-glycerol phosphate), stimulated with Dex (0, 1, 10 and 100 nM) based on previous literature report 21 (link), and with/without AKT inhibitor: LY294002 (10 μM, MedChemExpress, 154447-36-6). Medium was changed every 4 days. Alizarin red staining and Alizarin red absorbance at 562 nm were used to evaluate the degree of calcification.

Chen L., Ni Z., Huang J., Zhang R., Zhang J., Zhang B., Kuang L., Sun X., Zhang D., Su N., Qi H., Yang J., Jin M., Luo F., Chen H., Zhou S., Du X., Ouyang J., Wang Z., Xie Y., Tan Q, & Chen L. (2021). Long term usage of dexamethasone accelerating the initiation of osteoarthritis via enhancing the extracellular matrix calcification and apoptosis of chondrocytes. International Journal of Biological Sciences, 17(15), 4140-4153.

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Mouse Palate Culture and Transforming Growth Factor Signaling

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All animal care and experiments were performed under protocols approved by the Institutional Animal Care and Use Committees of the Baylor College of Dentistry and the University of Nebraska Medical Center. Mouse palate culture was performed as previously described (Kang and Svoboda, 2002 ; Yu et al., 2008 ; San Miguel et al., 2011 ). In brief: Palatal shelves were dissected from e13.5 CD1 mouse embryos and placed nasal side down on polycarbonate membranes (Nucleopore Corp.) with their medial edges in contact. The tissues were cultured with BGJb medium (Gibco) for 72 h. Medium was replaced every 24 h with fresh treatments. Anti-Tgfβ3 was used at concentration of 10 μM. TgfβrI Kinase Inhibitor VI (SB431542) was used at a concentration of 25 μM. Based on our initial dose-response experiments (not shown), this was the concentration of kinase inhibitor that abolished MES degradation in cultured palates while showing no signs of altered cell morphology. EphB2/Fc and control IgG Fc proteins were used at 5 μg/mL, as in our previously published studies. Fc proteins were clustered by mixing with anti-human Fc in a 4 to 1 w/w ratio and incubated at 22°C for 1 h or overnight at 4°C. This treatment allows the soluble Fc proteins to mimic the clustering that occurs on cell membranes and is required to initiate biologically relevant signaling.

SERRANO M.J., LIU J., SVOBODA K.K., NAWSHAD A, & BENSON M.D. (2015). Ephrin Reverse Signaling Mediates Palatal Fusion and Epithelial-to-Mesenchymal Transition Independently of Tgfβ3. Journal of cellular physiology, 230(12), 2961-2972.

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Murine Palatal Shelf Isolation and Culture

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On E14, wild-type C57BL/6J mouse embryos were quickly immersed in BGJb medium (Gibco). The palatal shelves were removed using forceps under a dissecting microscope. Isolated palatal shelves were placed in pairs on 0.4 μm porosity filters (Millipore, Burlington, MA, USA), nasal epithelium down, media edges in contact, on 35 mm tissue culture dishes (FALCON, France). The culture medium was composed of BGJb medium with or without Pan-Ras-IN-1 and/or SB431542. Pan-Ras-IN-1 was purchased from MedChemExpress (Monmouth Junction, NJ, USA). SB431542 was purchased from Sigma-Aldrich (St Louis, MO, USA). Samples were pretreated for 6 h prior to organ culture. Palatal shelves were cultured at 37°C with 5% CO₂. The culture medium and treatment solutions were replaced every 24 h.

Inubushi T., Fujiwara A., Hirose T., Aoyama G., Uchihashi T., Yoshida N., Shiraishi Y., Usami Y., Kurosaka H., Toyosawa S., Tanaka S., Watabe T., Kogo M, & Yamashiro T. (2022). Ras signaling and RREB1 are required for the dissociation of medial edge epithelial cells in murine palatogenesis. Disease Models & Mechanisms, 15(2), dmm049093.

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Mouse Embryonic Head Dissection

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The mouse embryonic heads were dissected in BGJb medium (Gibco). The palate was evaluated by direct observation and with a dissecting microscope. These tissue were fixed in 4% paraformaldehyde, equilibrated in graded sucrose, and embedded in Tissue-Tek (OCT compound, Sakura).

Sarper S.E., Kurosaka H., Inubushi T., Ono Minagi H., Kuremoto K.I., Sakai T., Taniuchi I, & Yamashiro T. (2018). Runx1-Stat3-Tgfb3 signaling network regulating the anterior palatal development. Scientific Reports, 8, 11208.

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Organ Culture and Radiolabeling of Tooth Germs

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An organ culture system was created according to a previous study.^{16 (link)} Based on the results of in situ hybridization/ immunohistochemistry, we selected the lower first molar tooth germs of E16.0, E18.0, and P3.0 as representatives of the late cap stage, predentin formation stage, and enamel and dentin formation stage, respectively. A total of 36 explants of tooth germs were cultured on cell culture inserts (Millicell, Merck, Tokyo, Japan) containing BGJB medium (Gibco: Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin (Gibco) for 2 h in a humidified atmosphere of 5% CO₂ in air at 37°C. Three tooth germs were placed on a cell culture insert. These explants were labeled with [³⁵S]Na₂SO₄ (3.7 MBq/mL, American Radiolabeled Chemicals, St Louis, MO, USA) for another 12 h under the same culture conditions.

Randilini A., Fujikawa K, & Shibata S. (2020). Expression, localization and synthesis of small leucine-rich proteoglycans in developing mouse molar tooth germ. European Journal of Histochemistry : EJH, 64(1), 3092.

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Osteoblast Phenotype and Signaling Analysis

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The subchondral bone plate was isolated and the cell cultures were prepared as previously described [5 (link)]. At confluence, cells were passaged once at 25000 cells/cm² and grown for 5 days in BGJb medium (Gibco) containing 10% fetal bovine serum (FBS). Confluent cells were then incubated in the presence or absence of 1,25-dihydroxyvitamin D₃ (1,25[OH]₂D₃; 50 nM) for 48 hours for the determination of biomarkers. Supernatants were collected at the end of the incubation. Cells were prepared in alkaline phosphatase buffer (100 mM glycine, 1 mM MgCl, 1 mM ZnCl, 1% Triton X-100; pH 10.5) for protein determination and phenotype evaluation, in TRIzol^™ for qRT-PCR experiments, or Laemmli buffer for Western blot analysis. Protein determination was performed by the bicinchoninic acid method. Wnt5a signaling activity was stimulated using recombinant human Wnt5a (rhWnt5a) protein at 100 ng/mL (R&D Systems) and TGF-β activity with recombinant human TGF-β1 (rhTGF-β1) at 10 ng/mL (R&D Systems). The expression of Wnt5a and TGF-β1 was inhibited in OA Ob by specific siRNA as previously described [5 (link)]. siWnt5a, siTGF-β1 and siScrambled (siSCR) preparations were Dharmacon SmartPOOL ONtarget products (a mix of 4 siRNA per target). For prolonged silencing treatments using siRNA in post-confluent osteoblasts, the treatments were repeated every 3 days of continuous culture.

Martineau X., Abed É., Martel-Pelletier J., Pelletier J.P, & Lajeunesse D. (2017). Alteration of Wnt5a expression and of the non-canonical Wnt/PCP and Wnt/PKC-Ca2+ pathways in human osteoarthritis osteoblasts. PLoS ONE, 12(8), e0180711.

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Palate Explant Culture with TGFβ3

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The palate was dissected and explanted from the E15.0 embryo and cultured on track-etched polycarbonate membrane filter (Nuclepore) in Trowell type organ culture with serumless, chemically defined BGJb medium (Gibco). In our dissection, the primary palate and the nasal septum were not excluded from our culture system. Affi-Gel beads (Bio-Rad) were incubated in TGFB3 (100 ng/μl, R&D Systems). Bovine serum albumin (BSA; Sigma-Aldrich) was used instead of recombinant protein for the control beads. The beads were immersed in recombinant protein or BSA at 37 °C for 60 min and placed on the primary palate of the explants using a pipette tube. After culture, the in vitro explants were fixed at each stage in 4% paraformaldehyde overnight and then processed for histological examination and qPCR analyses.

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Culturing Mouse Calvarias for Micro-CT Analysis

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Calvarias were cultured according to the literature.⁴³ (link) Briefly, calvarias were dissected from 5-day-old mice, then they were cut along the midline into two parts. The explants were subsequently cultured in BGJb medium (Gibco) containing 1 mg/mL BSA, 2 mM glutamine, and antibiotics supported by metal grids in a 6-well plate at 37°C in a humidified atmosphere of 5% CO₂ in air. The left parts of calvarias were treated with S2, and right parts were treated with a scrambled control siRNA. After 7 days of culture, the cultured calvarias were fixed with 4% paraformaldehyde (PFA) at 4°C, and then micro-CT scanning was taken for three-dimensional reconstruction. After being decalcified, dehydrated, and embedded in paraffin, coronal sutures were sectioned along the midline of the calvarias at 6-μm intervals and stained with H&E.

Luo F., Xie Y., Wang Z., Huang J., Tan Q., Sun X., Li F., Li C., Liu M., Zhang D., Xu M., Su N., Ni Z., Jiang W., Chang J., Chen H., Chen S., Xu X., Deng C., Wang Z., Du X, & Chen L. (2018). Adeno-Associated Virus-Mediated RNAi against Mutant Alleles Attenuates Abnormal Calvarial Phenotypes in an Apert Syndrome Mouse Model. Molecular Therapy. Nucleic Acids, 13, 291-302.

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Calvaria Bone Culture and Lumican Silencing

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The calvaria bone of timed-pregnant ICR mice (Orientbio) at E21.0 was cut in half along the calvaria sagittal line to include the frontal, coronal, and lambdoid sutures. The bone was placed in a transwell with an 8-μm-pore size polycarbonate membrane. To silence lumican expression, each calvaria bone was infected with lumican shRNA CM or control shRNA CM in BGjb medium (Gibco) containing 100 U/mL penicillin, and 100 μg/mL streptomycin and 0.1% BSA in the presence of 8 μg/mL polybrene. After 3 days, each bone was cultured for an additional 10 days in a medium containing 50 μg/mL of insulin (Sigma-Aldrich). The bone then was fixed in 4% PFA for 24 h and decalcified in 0.5 M EDTA in PBS for 2 days. The calvaria specimens were embedded in paraffin, coronally sectioned at a thickness of 3 μm, and stained with H&E or toluidine blue O solution (Marino et al., 2016 (link)). Calvaria bone widths and the number of osteoblasts were assessed using the same methods described above.

Lee J.Y., Park S.J., Kim D.A., Lee S.H., Koh J.M, & Kim B.J. (2020). Muscle-Derived Lumican Stimulates Bone Formation via Integrin α2β1 and the Downstream ERK Signal. Frontiers in Cell and Developmental Biology, 8, 565826.

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