Gridded scoring dish

Manufactured by STEMCELL

Sourced in Canada, Japan

The Gridded Scoring Dish is a laboratory equipment designed to assist with the handling and manipulation of cell cultures. It features a grid pattern to provide a visual reference for tracking and counting cells. The dish is made from durable materials suitable for cell culture applications.

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Lab products found in correlation

Eclipse te300, by Nikon (4 mentions) Methocult sfbit, by STEMCELL (3 mentions) Eclipse te3000, by Nikon (2 mentions) Anti il 10, by Abcam (1 mentions) Primaria dishes, by BD (1 mentions) 35 mm dishes, by BD (1 mentions) Methocult sf m3236, by STEMCELL (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Nichirei Biosciences (1 mentions)

8 protocols using gridded scoring dish

Angiogenic Potential of PBMNCs in MMD

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To determine the angiogenic potential of PBMNCs, cells derived from patients with MMD (n = 23), patients with MMD-O (n = 7), and controls (n = 23) were cultured in semisolid medium (MethoCult SFBIT; STEMCELL Technologies Inc., Vancouver, British Columbia, Canada) with provasculogenic growth factors/cytokines on 35-mm Primaria dishes (BD Falcon, USA) as reported previously^{10 (link),11 (link)}. EPC colonies that adhered to the dishes were then counted^{9 (link)}. Aliquots of the cells were seeded at 2 × 10⁵ cells/dish (three dishes per PBMNCs/cultured PBMNCs) for the EPC-CFA. The number of adherent colonies per dish was measured using a gridded scoring dish (STEM CELL Technologies, Canada) under a phase-contrast light microscope (Eclipse TE300; Nikon, Tokyo, Japan) 16 to 20 days after the initiation of the culture.
We measured the numbers of small- and large-cell colonies per dish after IL-10 (Pepro Tech, Inc., TX, USA) or anti-IL-10 (Abcam, UK) antibody was added to the freshly isolated PBMNCs or those cultured from patients with MMD (n = 7), patients with MMD-O (n = 3) and controls (n = 6).

Nagata E., Masuda H., Nakayama T., Netsu S., Yuzawa H., Fujii N., Kohara S., Sorimachi T., Osada T., Imazeki R., Matsumae M., Asahara T, & Takizawa S. (2019). Insufficient production of IL-10 from M2 macrophages impairs in vitro endothelial progenitor cell differentiation in patients with Moyamoya disease. Scientific Reports, 9, 16752.

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Neurocult NCFC Assay for Sphere Formation

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Cells in semi-solid media were prepared using the Neurocult NCFC Assay Kit (Stem Cell Technologies) per manufacturer’s protocol. Cells were plated at a density of 1650 cells/ml using 1.5 ml per 35 mm culture dish. Dishes were replenished after 7 days with 60 µl of neural stem cell proliferation media supplemented with heparin, EGF, and FGF. Sphere number and size were scored using a gridded scoring dish (Stem Cell Technologies). Eight-cell aggregates were used as the cutoff for scoring.

Ahmad S.T., Rogers A.D., Chen M.J., Dixit R., Adnani L., Frankiw L.S., Lawn S.O., Blough M.D., Alshehri M., Wu W., Marra M.A., Robbins S.M., Cairncross J.G., Schuurmans C, & Chan J.A. (2019). Capicua regulates neural stem cell proliferation and lineage specification through control of Ets factors. Nature Communications, 10, 2000.

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Evaluating CD34+ Cell Vasculogenic Potential

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To investigate the vasculogenic potential of nonexpanded and expanded CD34⁺ cells, we performed the EPC-CFU assay using semi-solid culture medium (MethoCult SF; StemCell Technologies) with proangiogenic growth factors in 35-mm Primaria dishes (BD Biosciences), as described previously.^{5 (link),22 (link),37–40} Twenty days after initiation of the culture, the number of adherent colonies per dish was measured using a gridded scoring dish (StemCell Technologies) under light microscopy. The primitive EPC-CFUs (small cells) and definitive EPC-CFUs (large cells) were counted separately.^{5 (link),22 (link),37–40}

Nakamura T., Koga H., Iwamoto H., Tsutsumi V., Imamura Y., Naitou M., Masuda A., Ikezono Y., Abe M., Wada F., Sakaue T., Ueno T., Ii M., Alev C., Kawamoto A., Asahara T, & Torimura T. (2016). Ex vivo expansion of circulating CD34+ cells enhances the regenerative effect on rat liver cirrhosis. Molecular Therapy. Methods & Clinical Development, 3, 16025-.

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Vasculogenic Potential of PBMNCs via EPC-CFA

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To investigate the vasculogenic potential of PBMNCs or QQMNCs, we used semisolid culture medium and 35‐mm Primaria dishes (BD Falcon; BD Biosciences) to grow and then counted the adhesive EPC colonies by EPC colony‐forming assay (EPC‐CFA) (MethoCult SF^BIT; STEMCELL Technologies Inc., Vancouver, BC, Canada) with proangiogenic growth factors/cytokines, as previously reported (Table 3).^{12 (link)} Aliquots of those cells were seeded at 2×10⁵ cells/dish (3 dishes per volunteer) for EPC‐CFA. Sixteen to 18 days after initiation of the culture, the number of adherent colonies per dish was measured using a gridded scoring dish (STEMCELL Technologies) under phase‐contrast light microscopy (Eclipse TE300; Nikon, Tokyo, Japan). Primitive EPC colony‐forming units (pEPC‐CFUs) and definitive EPC‐CFUs (dEPC‐CFUs) were separately counted.

Masuda H., Tanaka R., Fujimura S., Ishikawa M., Akimaru H., Shizuno T., Sato A., Okada Y., Iida Y., Itoh J., Itoh Y., Kamiguchi H., Kawamoto A, & Asahara T. (2014). Vasculogenic Conditioning of Peripheral Blood Mononuclear Cells Promotes Endothelial Progenitor Cell Expansion and Phenotype Transition of Anti‐Inflammatory Macrophage and T Lymphocyte to Cells With Regenerative Potential. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 3(3), e000743.

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Vasculogenic Potential of Cells

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To investigate vasculogenic potential, cells were cultured in semisolid medium (M3236 MethoCult SFBIT; Stemcell Technologies Inc., Vancouver, BC, Canada) with proangiogenic growth factors/cytokines, as previously reported, on 35-mm Primaria dishes (BD Falcon; BD Biosciences, USA) and then adhesive EPC colonies were counted. [13 (link)] Aliquots of the cells were seeded at 2 x 10⁵ cells/dish (3 dishes per PB-MNC/ RACs) for EPC-CFA. Ten to 12 days after initiation of culture, the number of adherent colonies per dish was measured using a gridded scoring dish (Stemcell Technologies, Vancouver, Canada) under a phase-contrast light microscope (Eclipse TE300; Nikon, Tokyo, Japan).

Nakayama T., Nagata E., Masuda H., Asahara T, & Takizawa S. (2019). Regeneration-associated cell transplantation contributes to tissue recovery in mice with acute ischemic stroke. PLoS ONE, 14(1), e0210198.

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Quantification of Primitive and Definitive EPC Colonies

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Freshly isolated PBMCs (1.5 × 10 5 ) were seeded into semisolid methyl cellulose-based culture medium coated 35 mm dishes (Falcon, Corning, CA, USA) and left in a humidified incubator with 5% CO2 at 37 °C until colony appearance. The number of adherent colonies was counted between days 16-18, using a gridded scoring dish (STEMCELL Tech., Vancouver, Canada) under a phase-contrast light microscope (Eclipse TE3000; Nikon, Tokyo, Japan). Primitive EPC colony-forming units (PEPC-CFUs) and definitive EPC colonyforming units (DEPC-CFUs) were separately counted as reported elsewhere 12, 13 .

Salybekov A.A., Sakai K., Natsumeda M., Vorateera K., Kobayashi S., Ikari Y., & Asahara T. (2020). Peripheral Levels of EPCs, T Cells, and Macrophages’ as Possible Predictive Biomarkers of Acute Myocardial Infarction.

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Enumeration of EPC Colonies from PBMNCs and BMMNCs

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Freshly isolated human or mouse peripheral blood mononuclear cells (PBMNCs), and mouse BM mononuclear cells (BMMNCs) were cultured in semisolid methyl cellulose-based culture medium, (MethoCult^™ SF M3236, STEMCELL Technologies Inc., Vancouver, BC, Canada) containing 100 ng/mL SCF, 50 ng/mL VEGF, 50 ng/mL basic fibroblast growth factor (bFGF), 50 ng/mL epidermal growth factor (EGF), 50 ng/mL insulin-like growth factor (IGF), 50 ng/mL interleukin-3 (IL-3) (these six proteins were purchased from Peprotech, Inc. Rocky Hill, NJ, USA), 2 IU/mL heparin (Ajinomoto Pharmaceutical Co. Ltd. Tokyo, Japan), 30% (v/v) fetal bovine serum (Nichirei Biosciences Inc., Tokyo, Japan) and penicillin/streptomycin (100 U/100 μg/mL; Gibco). Cells were seeded at 1.5 × 10⁵ cells/35 mm dish (BD Falcon, BD Bioscience, San Jose, CA, USA) and left in a humidified incubator with 5% CO₂ at 37°C till EPC colony formation. The number of adherent colonies on the dishes was counted between day 6–10 (mouse) and 16–18 (human) using gridded scoring dish (STEMCELL Technologies Inc. Vancouver, BC, Canada) under a phase-contrast light microscope (Eclipse TE3000; Nikon, Tokyo, Japan). Primitive EPC colony-forming units (pEPC-CFUs) and definitive EPC colony-forming units (dEPC-CFUs) were separately counted [16 (link)].

Salybekov A.A., Masuda H., Miyazaki K., Sheng Y., Sato A., Shizuno T., Iida Y., Okada Y, & Asahara T. (2019). Dipeptidyl dipeptidase-4 inhibitor recovered ischemia through an increase in vasculogenic endothelial progenitor cells and regeneration-associated cells in diet-induced obese mice. PLoS ONE, 14(3), e0205477.

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Quantifying Angiogenic Potential of Blood Cells

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The angiogenic potential of PBMNCs and QQMNCs was assessed using the EPC-CFA, as previously described^{6 (link),14 (link)}. The EPC-CFA was designed to differentiate total EPC colony-forming units (tEPC-CFUs) into two types of EPC-CFUs (primitive and definitive). Primitive EPC-CFU (pEPC-CFUs) are a predominantly proliferative population of cells, and definitive EPCs-CFU (dEPC-CFU) are a predominantly vasculogenic population with greater adhesion, migration, differentiation, and tubularization potential.
Briefly, adhesive EPC colonies were counted using the EPC-CFA (MethoCult SFBIT; Stem Cell Technologies Inc., Vancouver, BC, Canada) with semisolid culture medium containing proangiogenic growth factors/cytokines in 35-mm Primaria dishes (BD Falcon; BD Biosciences). Twelve aliquots of cells were seeded at 2 × 10⁵ cells/dish (three dishes per subject) for EPC-CFA. Sixteen to 18 days after culture initiation, the number of adherent colonies per dish was measured using a gridded scoring dish (Stem Cell Technologies) under phase-contrast light microscopy (Eclipse TE300; Nikon, Tokyo, Japan). The experiments were performed in triplicate. Experimental conditions were masked and two investigators counted the number of pEPC-CFUs and dEPC-CFUs.

Hagiwara H., Higashibata A., Ogawa S., Kanazawa S., Mizuno H, & Tanaka R. (2018). Effectiveness of endothelial progenitor cell culture under microgravity for improved angiogenic potential. Scientific Reports, 8, 14239.

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