35 mm culture dish

Manufactured by Corning

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The 35-mm culture dish is a laboratory equipment item used for culturing and maintaining cells in a controlled environment. It provides a suitable surface area and volume for cell growth and experimentation.

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35-mm dishes (42 citations) 35 mm culture dishes (21 citations) 35-mm dish (13 citations) 35-mm cell culture dish (11 citations) 35-mm polystyrene culture dish (2 citations)

26 protocols using 35 mm culture dish

Isolation and Culture of Rat Cerebrocortical Cells

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Mixed rat cerebrocortical cells containing neuronal and non-neuronal cells were separated from the brains of SD rat embryos at 17-days of gestation as described previously [29 (link),30 (link),31 (link)]. Briefly, cerebral cortices of the embryos were dissected and dissociated mechanically into single cells by trituration using fire-polished Pasteur pipettes. The isolated cells were plated in MEM supplemented with 5% FBS, 5% HS and 1% antibiotic-antimycotic agent at a density of 6 × 10⁵ cells/well of a 24-well culture plate (BD Falcon, Franklin Lakes, NJ, USA) or at a density of 6 × 10⁶ cells per 35 mm culture dish (Corning, NY, USA) pre-coated with laminin and poly-l-lysine. For immunocytochemical staining experiments, cells were plated at a density of 3 × 10⁵ cells per microscope cover glass (Marienfeld GmbH, Lauda-Königshofen, Germany), placed on each well of a 24-well culture plate. The cells were incubated at 37 °C in a humidified atmosphere of 95% air/5% CO₂. At 7 days after plating, proliferation of non-neuronal cells was arrested by the addition of 10 μM cytosine arabinoside. All experiments were conducted at 10–12 days after plating.

Lee K., Park C., Oh Y., Lee H, & Cho J. (2018). Antioxidant and Neuroprotective Effects of N-((3,4-Dihydro-2H-benzo[h]chromen-2-yl)methyl)-4-methoxyaniline in Primary Cultured Rat Cortical Cells: Involvement of ERK-CREB Signaling. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(3), 669.

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Isolation of Mouse Sperm and Oocytes

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Mice were euthanized by cervical dislocation. Both caudal epididymides were removed aseptically from the male mice and placed into 500 μL TYH buffer in a 35 mm culture dish (Corning, NY, USA). The epididymides were minced 5–6 times using forceps and scissors, and spermatozoa were allowed to disperse by gently shaking the dish by hand for 3–5 min at room temperature. Ovaries were collected from female mice and minced in TYH buffer; dispersed oocytes were collected using micropipettes.

Wang L., Chang S., Wang Z., Wang S., Huo J., Ding G., Li R., Liu C., Shangguan S., Lu X., Zhang T., Qiu Z, & Wu J. (2017). Altered GNAS imprinting due to folic acid deficiency contributes to poor embryo development and may lead to neural tube defects. Oncotarget, 8(67), 110797-110810.

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Islet Cell Aggregation Protocol

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Prior to functional readout and islet transplantation, the dissociated single islet cells were aggregated as cell clusters as previously described.¹⁷ Briefly, the dissociated islet cells were washed twice with HBSS after viral infection. The islet cells (2 × 10⁵ cells/dish) were then seeded into a non‐adhesive 35 mm culture dish (Corning) and cultured in RPMI‐1640 medium for 4 days until islet‐like cell clusters were formed.

Wang C., Du X., Fu F., Li X., Wang Z., Zhou Y., Gou L., Li W., Li J., Zhang J., Liao G., Li L., Han Y., Tong N., Liu J., Chen Y., Cheng J., Cao Q., Ilegems E., Lu Y., Zheng X, & Berggren P. (2022). Adiponectin gene therapy prevents islet loss after transplantation. Journal of Cellular and Molecular Medicine, 26(18), 4847-4858.

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Culturing Human Nasal Polyp Cells

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HNPCEC were purchased from ScienceCell Research Laboratories (Carlsbad, CA, USA) and cultured in Dulbecco’s modified Eagle Medium (DMEM) (Invitrogen, Grand Island, NY, USA) supplemented with 10% newborn calf serum (NCS; Invitrogen) and 100 units/ml penicillin and 100 µg/ml streptomycin (Roche Diagnostics, Mannheim, Germany) at 37°C in humidified air containing 5% CO₂. When outgrowth of the cells reached confluence, they were harvested with 0.025% trypsin (Invitrogen) in PBS, and transferred to plastic culture dishes at a 1:4 split ratio. For experiments, the cells were trypsinized and seeded at 1×10⁶ cells/ml per 35-mm culture dish (Corning Inc., Corning, NY, USA). The HNPCEC were found to be confluent after 72 hr (this day being set as day 0). Three different lots of HNPCEC were used (lot no. 0564, 0579, 5968), and employed in experiments from the 3rd to 6th passages in this study.

Fujita T., Tsuruga E., Yamanouchi K., Sawa Y, & Ishikawa H. (2014). Microfibril-Associated Glycoprotein-1 Controls Human Ciliary Zonule Development In Vitro. Acta Histochemica et Cytochemica, 47(1), 11-17.

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Establishment of Lens Epithelial Explants

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Gclc^w/w mice aged P20 were used for the establishment of lens epithelial explants, as previously described [35 , 36 (link)]. Briefly, mice were anesthetized, euthanized and their eyes enucleated as described in the histological analysis section (above). The lenses were removed and placed into a 35 mm culture dish (Corning, NY) containing pre-warmed (37°C) Medium 199 supplemented with 0.1% fetal bovine serum (Sigma-Aldrich, MO), 1% antibiotic-antimycotic (Sigma-Aldrich, St. Louis, MO) or Medium 199 supplemented with 0.1% fetal bovine serum (Sigma-Aldrich, MO), 1% antibiotic-antimycotic (Sigma-Aldrich, St. Louis, MO), and 10 mM NAC (Thermo Fisher Scientific). A hole was made at the posterior pole, the lens capsule opened, and the fiber cells gently removed. The lens capsule was pinned to the bottom of the culture dish such that the adherent epithelial cells were exposed to the medium. Explants were then individually cultured in a humidified atmosphere of 5% CO2 at 37°C for 24 hours. Six explants were pooled to make one sample and 3 pooled samples were used for each experimental condition (hence a total of 18 mice were used per condition).

Thompson B., Davidson E.A., Chen Y., Orlicky D.J., Thompson D.C, & Vasiliou V. (2022). Oxidative stress induces inflammation of lens cells and triggers immune surveillance of ocular tissues. Chemico-biological interactions, 355, 109804.

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Nanostructured Substrate Preparation for Cell Culture

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Custom-made sample dishes were prepared for experiments. A 15-mm cork borer was heated over an open flame to 100°C and then immediately pressed into the center of a 35-mm culture dish (Corning, Midland, MI). The edges of the hole were sanded down until smooth. The nanostructured PMMA-on-glass coverslips were mounted to the bottom of the dishes with clear silicone adhesive (Corning, Midland, MI) so that the nanostructured region was placed in the center of the opening. The mounted samples were dried overnight before use. At the time of cell culture, the samples were UV-sterilized for 5 minutes. Nanostructured surfaces were coated with 1 μg/mL of fibronectin (Sigma-Aldrich, St. Louis, MO) for 1 hour at 37°C before cell seeding.

Liang E.I., Mah E.J., Yee A.F, & Digman M.A. (2017). Correlation of Focal Adhesion Assembly and Disassembly with Cell Migration on Nanotopography. Integrative biology : quantitative biosciences from nano to macro, 9(2), 145-155.

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Quantifying Cell Migration via Wound Assay

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Cell migration was measured as described previously, with minor modifications (Ryu et al., 2012 (link)). Briefly, when MH7A cells reached confluence in a 35-mm culture dish (Corning, NY, USA), three wound lines in the form of a cross were made by scratching the cellular layer with a plastic pipette tip. Floating cells were then washed out, and fresh medium was added. Cells were then incubated under normoxia condition. Narrowing of the wound was then monitored using a phase-contrast microscope beginning 6 h after the scratch. Cells were fixed and stained in a 20% methanol/0.1% crystal violet solution during 1 h at 4°C temperature, followed by washing the cells with water to remove redundant staining. The size of the wound at each time point was then quantified using NIH image analysis software (Image J, version 1.62; National Institutes of Health, MD, USA), and compared with that in the initiation of cell migration.

Lee J., Yoon S.S., Thuy P.X, & Moon E.Y. (2020). Synovial Cell Migration is Associated with B Cell Activating Factor Expression Increased by TNFα or Decreased by KR33426. Biomolecules & Therapeutics, 28(5), 405-413.

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Retinal Explant Culture Procedure

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The experimental procedures were approved by Institutional Animal Care and Use Committee of Tianjin Medical University (Permit Number: SYXK 2009-0001) and complied with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. Fertilized eggs were incubated at 38 °C and 60% relative humidity. The retinal explant cultures were set up as previously described with modifications31 (link)55 (link). On embryonic day 9 (E9), the retina was isolated and cultured on an organotypic insert (0.4 μm pore size, Merk Millipore, Billerica, MA, USA) in a 35-mm culture dish (Corning, Corning, NY, USA) with photoreceptor facing down. The explants were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM, 4.5 g/L glucose, Life Technologies, Grand Island, NY, USA) supplemented with 10% or 15% Fetal Bovine Serum (FBS, Life Technologies, Grand Island, NY, USA), 100 U/ml penicillin/100 μg/ml streptomycin (Life Technologies, Grand Island, NY, USA), and 2 mM L-glutamine (Life Technologies, Grand Island, NY, USA) at 37 °C and 5% CO₂. The culture media were changed every other day.

Zhang Y., Bo Q., Wu W., Xu C., Yu G., Ma S., Yang Q., Cao Y., Han Q., Ru Y., Liu X., Hua Wei R., Wang F.E., Zhang X, & Li X. (2015). α-Melanocyte-stimulating hormone prevents glutamate excitotoxicity in developing chicken retina via MC4R-mediated down-regulation of microRNA-194. Scientific Reports, 5, 15812.

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Adipogenic Differentiation of Third-Passage Cells

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The third-passage cells were seeded at a density of 1 × 10⁵/ml in a 35-mm culture dish (Corning). When the cell confluence reached approximately 90%, the medium was replaced with an adipogenic medium (α-MEM medium supplemented with 10% FBS, 100-U/ml penicillin, 100-μg/ml streptomycin, 1% L-glutamine, 1 μM dexamethasone, 0.2 mM indomethacin, 0.5 mM 3-isobutyl-methylxanthine, 10-µg/ml insulin [Sigma-Aldrich]). After induction for 21 days, the cells were stained with Oil Red O (Solarbio). The mRNA expression level of peroxisome proliferator-activated receptor gamma (PPAR-γ) was measured with real-time PCR.

Zhang P.P., Liang S.X., Wang H.L., Yang K., Nie S.C., Zhang T.M., Tian Y.Y., Xu Z.Y., Chen W, & Yan Y.B. (2021). Differences in the biological properties of mesenchymal stromal cells from traumatic temporomandibular joint fibrous and bony ankylosis: a comparative study. Animal Cells and Systems, 25(5), 296-311.

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Scratch Wound Assay for Cell Migration

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Cells (2 × 10⁶) in serum-free media were seeded on each 35-mm culture dish (Corning Incorporated). A pipette tip (0.1–10 μl, Labcon North America, Petaluma, CA, USA) was used to create a scratch wound in eight culture dishes. They were respectively incubated at AP (n = 4) and NP (n = 4) for 12 h. The scratched wound size was measured immediately and 12 h after wounding in the two different pressures. The difference between the initial distance and that after 12 h of incubation was defined as the migration distance [Fig. 1].Fig. 1

The difference in the scratched wound distance between the status immediately post wounding (0 h) and that after 12 h of incubation at the two different pressures was defined as the migration distance (n = 4 in each pressure condition). A significantly greater migration distance was observed in cells at negative-pressure for 12 h.

Chow S.E., Chen C.P., Hsu C.C., Tsai W.C., Wang J.S, & Hsu N.C. (2016). Quantifying cell behaviors in negative-pressure induced monolayer cell movement. Biomedical Journal, 39(1), 50-59.

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