Culture plates

Manufactured by Corning

Sourced in United States, Israel, China

Culture plates are a type of laboratory equipment used for the growth and cultivation of microbial or cell cultures. They provide a controlled and sterile environment for the growth of organisms, enabling researchers to study their properties and behavior.

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25 protocols using culture plates

Retinal Organoid Generation from hiPSCs

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Two human-induced pluripotent stem cell (hiPSC) lines, the BC1-GFP cell line (gifted by Prof. Linzhao Cheng, Johns Hopkins University School of Medicine) and the SB cell line (CA4002106, Cellapy, China), were used to generate retinal organoids. The hiPSCs were maintained in a feeder-free system in mTeSR1 medium (STEMCELL Technologies, Canada). They were then induced to differentiate into retinal organoids using the protocol published by Zhong et al. and Luo et al. [14 (link), 15 (link)]. Briefly, hiPSCs were maintained in suspension until they formed embryoid bodies (EBs), following which, the EBs were induced to form retinal organoids after being sequentially cultured in N2 (17502-048, Gibco) and B27 (12587-010, Gibco) supplements.
After 30-45 days of induction, some retinal organoids were harvested and fixed in 4% paraformaldehyde (Sigma-Aldrich) for 30 minutes, then dehydrated using graded sucrose (Sigma-Aldrich) and embedded in optimal cutting temperature compound (O.C.T., SAKURA, Japan). And retinal organoids were frozen and sectioned via cryotomy (Leica, Germany). Some retinal organoids were attached on coverslips (Jieli Biotechnology Co. Ltd., China) or culture plates (Corning) coated with Matrigel (354277, Corning) and cultured with either RAP (20 μg/ml) or DEX (170 μg/ml).

Xian B., Luo Z., Li K., Li K., Tang M., Yang R., Lu S., Zhang H, & Ge J. (2019). Dexamethasone Provides Effective Immunosuppression for Improved Survival of Retinal Organoids after Epiretinal Transplantation. Stem Cells International, 2019, 7148032.

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Osteoclast Differentiation Protocol

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Minimum Essential Medium α (Alpha-MEM) and fetal bovine serum (FBS), referred to here as “Standard medium,” were purchased from Rhenium (Modiin, Israel), and culture plates were from Corning (New York, NY, United States). As a source of M-CSF, we used supernatant from CMG 14–12 cells, containing 1.3μg/ml M-CSF (Takeshita et al., 2000 (link)). RANKL was purchased from R&D Systems, Minneapolis, MN, United States.

Cohen-Karlik E., Awida Z., Bergman A., Eshed S., Nestor O., Kadashev M., Yosef S.B., Saed H., Mansour Y., Globerson A., Neumann D, & Gabet Y. (2021). Quantification of Osteoclasts in Culture, Powered by Machine Learning. Frontiers in Cell and Developmental Biology, 9, 674710.

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Protocols for Ivermectin Experiments

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IVM (IVM), dimethylsulfoxide (DMSO), sodium hypochlorite, cholesterol, tetracyclin, ampicillin, isopropyl-β-D-thiogalactoside (IPTG), serotonin (5-hydroxytryptamine, 5HT), rhodamine 123 and fluorescein 5-isothiocyanate (FITC) were purchased from Aldrich (Sigma, Aldrich Chimie, St Quentin Fallavier, France). Moxidectin (MOX) was a generous gift from Fort Dodge International (Fort Dodge, IA). Culture plates were supplied by Corning (Borre, France). For all experiments, IVM was dissolved in DMSO and the maximal concentration of DMSO was 0.5% in all assays.

Ménez C., Alberich M., Courtot E., Guegnard F., Blanchard A., Aguilaniu H, & Lespine A. (2019). The transcription factor NHR-8: A new target to increase ivermectin efficacy in nematodes. PLoS Pathogens, 15(2), e1007598.

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3D Cell Culture on Matrigel

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Culture plates (35 mm; Corning Inc.) were coated with 100 µL of growth factor-reduced Matrigel (BD Biosciences) and were left to solidify for 30 min at 37°C. The cells were then plated at a concentration of 10⁴ cells/mL and cultured in normal or macrophage-conditioned medium. Cell growth on Matrigels was observed everyday using a phase contrast microscope.

Król M., Mucha J., Majchrzak K., Homa A., Bulkowska M., Majewska A., Gajewska M., Pietrzak M., Perszko M., Romanowska K., Pawłowski K., Manuali E., Hellmen E, & Motyl T. (2014). Macrophages Mediate a Switch between Canonical and Non-Canonical Wnt Pathways in Canine Mammary Tumors. PLoS ONE, 9(1), e83995.

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Macrophage Differentiation and Activation

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Alpha-MEM and fetal bovine serum (FBS), were purchased from Rhenium (Modiin, Israel) and culture plates were from Corning (New York, NY, USA). LPS from the E. coli strain 0127:B8 (Sigma) was reconstituted in sterile double distilled water (DDW) as a 1 mg/mL stock solution and kept at −20 °C. Dulbecco’s Phosphate Buffered Saline (DPBS) was purchased from Biological Industries. As a source of M-CSF, we used supernatant from CMG 14-12 cells containing 1.3 μg/mL M-CSF [38 (link),76 (link)]. RANKL was purchased from R&D Systems, Minneapolis, MN, USA. The peptide Cibinetide was kindly provided by Araim Pharmaceuticals, Inc. (Ossining, NY, USA). Erythropoietin (EPO) was obtained from GMP-manufactured sterile syringes containing rHuEPO (Epoetin alfa, Eprex^®) as used for patient care. These were kindly provided by Janssen Cilag, Israel, and were employed throughout this study.

Awida Z., Bachar A., Saed H., Gorodov A., Ben-Califa N., Ibrahim M., Kolomansky A., Iden J.A., Graniewitz Visacovsky L., Liron T., Hiram-Bab S., Brines M., Gabet Y, & Neumann D. (2021). The Non-Erythropoietic EPO Analogue Cibinetide Inhibits Osteoclastogenesis In Vitro and Increases Bone Mineral Density in Mice. International Journal of Molecular Sciences, 23(1), 55.

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Isolation and Culture of Murine Bone Marrow-Derived Mesenchymal Stem Cells

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The bone marrow of 14-day-old C57BL/6 mice was harvested and passed through a 40-μm cell strainer, yielding single cells. Subsequently, the single-cell suspension was incubated with antibodies specific for CD51-PE (1:200 dilution, BD, USA), CD45-FITC (1:200 dilution, eBioscience, USA), Ter119-PECY7 (1:200 dilution; eBioscience) and CD31-APC (dilution 1:200; eBioscience) at 4 °C for 30 min. CD51⁻CD45⁻Ter119⁻CD31⁻ cells (CD51⁻bMSCs) and CD51⁺CD45⁻Ter119⁻CD31⁻ cells (CD51⁺bMSCs) were sorted using flow cytometry (Influx, BD). Then, the isolated cells were allowed to adhere to culture plates (Corning, USA) in medium described as follows: DMEM/F12 (Gibco, USA) containing 10 ng/ml EGF (Pepro tech, USA), 10 ng/ml bFGF (Pepro tech), 2% B27 (Invitrogen, USA), 0.1 mM β-mercaptoethanol (Invitrogen), 1% l-glutamine (Sigma Aldrich, USA), 1% foetal bovine serum (Gibco) and 100 IU/ml penicillin/streptomycin (Invitrogen). Cells were cultured at 37 °C in a 5% CO₂ atmosphere and propagated every 2 or 3 days.

Xie D.M., Li Y.L., Li J., Li Q., Lu G., Zhai Y., Zhang J., Huang Z, & Gao X. (2019). CD51 distinguishes a subpopulation of bone marrow mesenchymal stem cells with distinct migratory potential: a novel cell-based strategy to treat acute myocardial infarction in mice. Stem Cell Research & Therapy, 10, 331.

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Differentiation of Osteoclast Precursors

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Alpha-MEM and fetal bovine serum (FBS) were purchased from Rhenium (Modiin, Israel), culture plates were from Corning (New York, NY, USA). As a source of M-CSF, we used supernatant from CMG 14-12 cells, containing 1.3 μg/mL M-CSF [18 (link),71 (link)]. RANKL was purchased from R&D Systems, Minneapolis, MN, USA. Erythropoietin (EPO) used in the study was obtained from GMP-manufactured sterile syringes containing rHuEPO (Epoetin alfa, Eprex^®, Janssen) as used for patient care. These were kindly provided by Janssen Cilag, Israel.

Awida Z., Hiram-Bab S., Bachar A., Saed H., Zyc D., Gorodov A., Ben-Califa N., Omari S., Omar J., Younis L., Iden J.A., Graniewitz Visacovsky L., Gluzman I., Liron T., Raphael-Mizrahi B., Kolomansky A., Rauner M., Wielockx B., Gabet Y, & Neumann D. (2022). Erythropoietin Receptor (EPOR) Signaling in the Osteoclast Lineage Contributes to EPO-Induced Bone Loss in Mice. International Journal of Molecular Sciences, 23(19), 12051.

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C2C12 Myoblast Differentiation Protocol

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C2C12 myoblasts (ATCC^® Number CRL-1772) were grown in high-glucose DMEM supplemented with 10% fetal calf serum (proliferation medium). When cells reached 70–80% confluence myoblasts were induced to differentiate and fuse into multinucleated myotubes by replacing the proliferation medium to DMEM with 2% donor horse serum (differentiation medium). After 6 days, multinucleated myotubes were infected for in vitro gene transfer protocol as described below. Chemicals were from Gibco^® (Thermofisher, Carlsbad, CA, USA) and Sigma-Aldrich^® (St Louis, MO, USA) and culture plates from Corning^® (New York City, NY, USA).

Pereyra A.S., Mykhaylyk O., Lockhart E.F., Taylor J.R., Delbono O., Goya R.G., Plank C, & Hereñu C.B. (2016). Magnetofection Enhances Adenoviral Vector-based Gene Delivery in Skeletal Muscle Cells. Journal of nanomedicine & nanotechnology, 7(2), 364.

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Analyzing Cell Growth on Matrigel

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Culture plates (35 mm; Corning Inc.) were coated with 100 µL of growth factor-reduced Matrigel (BD Biosciences) and were left to solidify for 30 min at 37°C. The control cells, siRNA-treated cells or IL-28 treated cells were then plated at a concentration of 10⁴ cells/mL and cultured for 24 hrs. Cell growth on Matrigels was observed using a phase contrast microscope.

Mucha J., Majchrzak K., Taciak B., Hellmén E, & Król M. (2014). MDSCs Mediate Angiogenesis and Predispose Canine Mammary Tumor Cells for Metastasis via IL-28/IL-28RA (IFN-λ) Signaling. PLoS ONE, 9(7), e103249.

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Spheroid Formation from Adherent Cell Lines

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Lids of 10 cm culture plates (Corning, NY) were seeded with 30 μl growth media containing 5,000 cells of either A375, BxPC3, PANC1 or MDA-MB-231. To prevent dehydration of the drops, 5 ml PBS were added to the bottom of the plate. Cells were incubated at 37 °C and 5% CO₂ for 48 hours to allow aggregation. Once cells created aggregates in the drops, spheroids were collected and transferred to a 10 cm culture plate coated with 0.75% agarose in 10 ml growth medium for an additional 5 days.

Shoval H., Karsch-Bluman A., Brill-Karniely Y., Stern T., Zamir G., Hubert A, & Benny O. (2017). Tumor cells and their crosstalk with endothelial cells in 3D spheroids. Scientific Reports, 7, 10428.

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