Culture insert

Manufactured by Merck Group

The Culture Insert is a laboratory equipment designed for cell culture applications. It provides a stable platform for cells to grow and proliferate in vitro. The insert features a porous membrane that allows for nutrient and gas exchange, supporting the maintenance of cell cultures.

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Lab products found in correlation

Phenol red free dmem, by Corning (1 mentions) B27 supplement, by Thermo Fisher Scientific (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Millicell Hanging Cell Culture Inserts (91 citations) Cell culture inserts (67 citations) Millicell culture plate inserts (27 citations) Tissue culture inserts (10 citations) 24-well plate cell culture insert (5 citations) Millicell 24-well culture insert plates (4 citations) Millicell-24 Cell Culture Insert Plate system (3 citations) Millicell Hanging Cell Culture Insert of 8 μm porosity (2 citations) Millicell Cell Culture Insert with 8-μM-pore Polyvinylpyrrolidone-free polycarbonate membranes (2 citations) Millicell tissue culture insert (2 citations)

2 protocols using culture insert

Dual-Plasmid Interneuron Electroporation

Cited 1 time

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A detailed protocol is described in our recent publication (Keener et al., 2020a (link); Keener et al., 2020b (link)). Briefly, the slice cultures were perfused with filter-sterilized aCSF consisting of (in mM): 119 NaCl, 2.5 KCl, 0.5 CaCl₂, 5 MgCl₂, 26 NaHCO₃, 1 NaH₂PO₄, 11 glucose, and 0.001 mM tetrodotoxin (TTX, Hello Bio Inc), gassed with 5% CO₂/95% O₂, pH 7.4. Patch pipettes (4.5–8.0 MΩ) were each filled with plasmids containing either tag-BFP and Nrxn or EGFP and Nlgn3Δ (0.05 µg/µl for each plasmid) and respectively electroporated in TdTomato-positive VGT3+ interneurons and CA1 pyramidal neurons. The same internal solution for single-cell sequencing was used. A single electrical pulse train (amplitude: −5 V, square pulse, train: 500 ms, frequency: 50 Hz, pulse width: 500 µs) was applied to the target neurons. After electroporation, the electrode was gently retracted from the cell and the slices were transferred to a culture insert (Millipore) with slice culture medium in a petri dish and incubated in a 5% CO₂ incubator at 35°C for 3 days.

Uchigashima M., Konno K., Demchak E., Cheung A., Watanabe T., Keener D.G., Abe M., Le T., Sakimura K., Sasaoka T., Uemura T., Imamura Kawasawa Y., Watanabe M, & Futai K. (2020). Specific Neuroligin3–αNeurexin1 signaling regulates GABAergic synaptic function in mouse hippocampus. eLife, 9, e59545.

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Culturing Mouse Circadian Tissues

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Following procedures reported previously (Buhr et al., 2010 (link); Izumo et al., 2014 ; Lee et al., 2015 (link)), the mouse brain was collected following cervical dislocation, before the onset of darkness. Peripheral tissues including liver were also collected. Coronal brain slices of 250–300 μm were made with a Vibratome in cold HBSS buffer. The SCN region was trimmed into a 1 mm × 1 mm square with optic chiasm attached and placed on top of a culture insert with pore size of 0.4 μm (Millipore). A 1 mm × 2 mm piece of liver was cultured in the same way as a control sample. All tissue slices were cultured in phenol red-free DMEM (Corning) supplemented with high glucose, glutamine, pyruvate, HEPES, and B27 supplement (Gibco). Culture medium was changed once a week. TTX treatment followed a previous report (Yamaguchi et al., 2003 (link)), except that 1 μM was applied for a period of a week. TTX was added by changing pre-mixed fresh medium, and wash-out of TTX was carried out by moving the culture insert to a new culture dish with pre-warmed fresh culture medium, four times, repeatedly.

Shan Y., Abel J.H., Li Y., Izumo M., Cox K.H., Jeong B., Yoo S.H., Olson D.P., Doyle FJ I.I.I, & Takahashi J.S. (2020). Dual-Color Single-Cell Imaging of the Suprachiasmatic Nucleus Reveals a Circadian Role in Network Synchrony. Neuron, 108(1), 164-179.e7.

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