35 mm culture dishes

Manufactured by Sarstedt

Sourced in United States, France

The 35 mm culture dishes are sterile plastic containers used for culturing cells and tissues in laboratory settings. They provide a controlled environment for the growth and maintenance of cell cultures.

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Lab products found in correlation

75 cm2 plastic culture flasks, by Thermo Fisher Scientific (4 mentions) Jetprime reagent, by Polyplus Transfection (3 mentions) Model ix71, by Olympus (3 mentions) Hek293t cell, by American Type Culture Collection (3 mentions) 3294 cellbind surface, by Corning (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Fetal calf serum (fcs), by American Type Culture Collection (1 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Hek293, by American Type Culture Collection (1 mentions) Muse cell cycle kit, by Merck Group (1 mentions) Laminin, by BD (1 mentions) Collagenase type 2, by Merck Group (1 mentions) Poly d lysine, by Merck Group (1 mentions) Dispase, by Merck Group (1 mentions)

6 protocols using 35 mm culture dishes

HEK293T Cell Transfection Protocol

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Experiments were performed using HEK293T cells (ATCC, Manassas, VA, USA), which were grown in Dulbecco’s modified Eagle’s medium (Gibco, Rockville, MD, USA) supplemented with 10% fetal calf serum (ATCC, Manassas, VA, USA), 50 U/mL penicillin, and 50 μg/mL streptomycin (Sigma, St Louis, MO, USA) in a humidified 5% CO₂ atmosphere at 37 °C. The cells were cultured in 75 cm² plastic culture flasks (NUNC, Rochester, NY, USA) for 36–72 h until they reached 80–95% confluence. On the day before transfection, ~150,000 cells were seeded on 35 mm culture dishes (Sarstedt, Newton, NC, USA) and incubated at 37 °C for at least 24 h. Transfection was performed using 2 μg of DNA and 2 μL of jetPRIME reagent in 2 mL of Dulbecco’s modified Eagle’s medium according to the manufacturer’s instructions (PolyPlus-transfection, Illkirch, France). After 24 h of incubation, the transfected cells were mechanically dispersed and reseeded on 35 mm Corning 3294 CellBIND Surface cell culture dishes (Corning, City of Corning, NY, USA) for 2–8 h prior to recording. For the dye uptake measurements, the transfected cells were plated on 12 mm poly-L-lysine-coated coverslips (Glaswarenfabrik Karl Hecht KG, Sandheim, Germany).

Rupert M., Bhattacharya A., Stillerova V.T., Jindrichova M., Mokdad A., Boué-Grabot E, & Zemkova H. (2020). Role of Conserved Residues and F322 in the Extracellular Vestibule of the Rat P2X7 Receptor in Its Expression, Function and Dye Uptake Ability. International Journal of Molecular Sciences, 21(22), 8446.

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Transfection of HEK-293T cells for P2Y receptor studies

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Experiments were performed on human embryonic kidney 293T cells (hereafter HEK-293T cells; American Type Culture Collection, Rockville, MD, USA) grown in Dulbecco modified Eagle's medium supplemented with 10% fetal bovine serum, 50 U/ml penicillin and 50 μg/ml streptomycin in a humidified 5% CO₂ atmosphere at 37 °C. These cells express endogenously several subtypes of metabotropic P2Y receptors [29 (link),8 (link),6 (link)], but not P2X4R [38 (link)]. Cells were cultured in 75 cm² plastic culture flasks (NUNC, Rochester, NY) for 36–72 h, until they reached 80–95% confluence. Before the day of transfection, ~150,000 cells were plated on 35 mm culture dishes (Sarstedt, Newton, NC) and incubated at 37 °C for at least 24 h. For each culture dish of HEK-293T cells, transfection of either wild type or mutant receptors was conducted using 2 μg of DNA with 2 μl of jetPRIME™ reagent in 2 ml of Dulbecco modified Eagle's medium, according to the manufacturer's instructions (PolyPlus-transfection, Illkirch, France). After 24–48 h of incubation, the transfected cells were mechanically dispersed and re-cultured on 35 mm dishes of Corning 3294 CellBIND Surface for 2 – 8 hours before recording. Transfected cells were identified by the fluorescence signal of GFP using an inverted research microscope with fluorescence illuminators (Model IX71; Olympus, Melville, NY).

Zemkova H., Khadra A., Rokic M.B., Tvrdonova V., Sherman A, & Stojilkovic S.S. (2014). Allosteric Regulation of the P2X4 Receptor Channel Pore Dilation. Pflugers Archiv : European journal of physiology, 467(4), 713-726.

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Recombinant Channel Expression in HEK 293T Cells

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To express the recombinant channels, we used human embryonic kidney (HEK) 293T cells (American Type Culture Collection, Rockville, MD, USA) grown in Dulbecco modified Eagle’s medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St Louis, MO), 50 U/ml penicillin and 50 µg/ml streptomycin (both Thermo Fisher Scientific, Waltham, MA) in a humidified 5% CO₂ and 95% air at 37°C. Cells were cultured in 75 cm² plastic culture flasks (NUNC, Rochester, NY) for 36–72 hours until reaching 80–95% confluence. Before the day of transfection, the cells were plated on 35 mm culture dishes (Sarstedt, Newton, NC) and incubated at 37°C for at least 24 h. Transfection was done using 2 µg of either WT or mutant receptor DNA with 2 µl of JetPrime reagent in 2 ml of Dulbecco modified Eagle’s medium, according to manufacturer’s instructions (PolyPlus-transfection, Illkirch, France). Transfected cells were identified by the fluorescence signal of EGFP using the Olympus IX71 inverted research microscope with fluorescence illuminators (Model IX71; Olympus, Melville, NY).

Tvrdonova V., Rokic M.B., Stojilkovic S.S, & Zemkova H. (2014). Identification of Functionally Important Residues of the Rat P2X4 Receptor by Alanine Scanning Mutagenesis of the Dorsal Fin and Left Flipper Domains. PLoS ONE, 9(11), e112902.

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Transient Transfection of HEK293 Cells

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Experiments were performed on human embryonic kidney 293 cells (HEK293; American Type Culture Collection), which were grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum, 50 U/mL penicillin, and 50 μg/mL streptomycin in a humidified 5% CO₂ atmosphere at 37°C. Cells were cultured in 75-cm² plastic culture flasks (NUNC, Rochester, NY, USA) for 36–72 h until they reached 80–95% confluence. Before the day of transfection, ~150 000 cells were plated on 35 mm culture dishes (Sarstedt, Newton, NC, USA) and incubated at 37°C for at least 24 h. For each culture dish of HEK293 cells, transfection of wild-type P2X4R was conducted using 2 μg of DNA with 2 μl of jetPRIME reagent in 2 ml of Dulbecco modified Eagle’s medium, according to the manufacturer’s instructions (PolyPlus-transfection, Illkirch, France). After 24–48 h of incubation, the transfected cells were mechanically dispersed and re-cultured on 35 mm dishes of Corning 3294 CellBIND Surface for 1–4 hours before recording. Transfected cells were identified by the fluorescence signal of enhanced green fluorescent protein using an inverted research microscope with fluorescence illuminators (Model IX71; Olympus, Melville, NY).

Mackay L., Zemkova H., Stojilkovic S.S., Sherman A, & Khadra A. (2017). Deciphering the regulation of P2X4 receptor channel gating by ivermectin using Markov models. PLoS Computational Biology, 13(7), e1005643.

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Ouabain-Induced Cell Cycle Analysis

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As described in our previous work [19 (link)], 5 × 10⁵ cells were seeded in 35 mm culture dishes (Sarstedt) in a final volume of 2 mL. Following 24 h of incubation in an atmosphere of 5% CO₂ at 37 °C, culture supernatants were removed and replaced with fresh culture medium (control) or culture medium plus 10⁻⁷ M of ouabain, and cells were cultivated for an additional 24 h. Cell cycle analysis was then performed according to the manufacturer’s instructions using a Muse Cell Cycle Kit (Millipore).

Teixeira M.P., Haddad N.F., Passos E.F., Andrade M.N., Campos M.L., da Silva J.M., de Figueiredo C.S., Giestal-de-Araujo E., de Carvalho D.P., Miranda-Alves L, & de Paiva L.S. (2022). Ouabain Effects on Human Anaplastic Thyroid Carcinoma 8505C Cells. Cancers, 14(24), 6168.

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Dissociation and Culture of DRG Neurons

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DRGs were extracted from adult TRPV1-ChR2 or MrgD-ChR2 mice and kept in sterile ice-cold 1x HBSS Hanks medium (Invitrogen) throughout the dissection. DRGs were then incubated in 5 mL of HBSS containing 1.4 mg/mL dispase (Sigma-Aldrich) and 1.1 mg/mL collagenase type II (Sigma-Aldrich) for 45 min at 37 o C. Following the enzymatic reaction, DRGs were washed once with F-12 culture media (Invitrogen), containing 10% FBS, 1% L-Glutamine, 1% penicillin, and 1% streptomycin, and then mechanically triturated using fire-polished Pasteur pipettes. The dissociated neurons were finally plated onto 35 mm culture dishes (Sarstedt, 2 mL/dish) previously coated with laminin (BD Bioscience) and poly-D-lysine (Sigma-Aldrich).
Cells were incubated for 2 h at 37 o C and 5% CO 2 prior to electrophysiological recording. Since a significant downregulation of capsaicin responses was observed in TRPV1-ChR2 neurons 24 to 72 h after plating (data not shown), electrophysiological recordings were conducted using freshly dissociated DRG neurons.

Beaudry H., Daou I., Ase A.R., Ribeiro-da-Silva A, & Séguéla P. (2017). Distinct behavioral responses evoked by selective optogenetic stimulation of the major TRPV1+ and MrgD+ subsets of C-fibers. Pain, 158(12).

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