100 μm cell strainer

Manufactured by Avantor

Sourced in United States, United Kingdom

The 100 μm cell strainer is a laboratory filtration device designed to separate cells or particles based on size. It features a mesh with 100 micrometer pore size to retain larger particles while allowing smaller components to pass through.

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Cell strainer

6 protocols using 100 μm cell strainer

Spleen Processing for Flow Cytometry Analysis

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The spleen was processed freshly as described previously with minor modifications (n = 9 CT26 injected, n = 9 vehicle injected control) [72 (link)]. Briefly, after the spleen was smashed on a 100 μm cell strainer, (VWR, Radnoe, PA, USA), washed with PBS, and centrifuged at 1400 rpm for 5 min, the pellet was resuspended in sterile PBS. Red blood cell lysis was carried out by the incubation of cells with 5 mL ACK solution for 5 min. Samples were loaded on a cell strainer (70 μm in pore size) and washed twice with 20 mL PBS. After that, cells were resuspended and pipetted into 12 × 75 mm FACS tubes (VWR International, PA, USA) and diluted with 100 µL staining buffer (PBS with 1% FBS, GIBCO, Life Technologies, Paisley, UK, and 0.1% sodium azide, Sigma-Aldrich, Saint Louis, MO, USA).

Faragó A., Zvara Á., Tiszlavicz L., Hunyadi-Gulyás É., Darula Z., Hegedűs Z., Szabó E., Surguta S.E., Tóvári J., Puskás L.G, & Szebeni G.J. (2024). Lectin-Based Immunophenotyping and Whole Proteomic Profiling of CT-26 Colon Carcinoma Murine Model. International Journal of Molecular Sciences, 25(7), 4022.

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Generation of Human Intestinal Organoids

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Human organoids were generated from biopsy samples collected at Weill Cornell Medicine or obtained from the In Vivo Animal and Human Studies Core at University of Michigan Center for Gastrointestinal Research (Key Resource Table). To generate organoids, human colon or ileum biopsy samples were cut into ~1 mm pieces and washed with cold DPBS by pipetting 2–3 times. Samples were treated with collagenase type IV (Worthington, 2 mg/ml in F12K medium) at 37°C for 30 mins with pipetting every 10 mins. Digestion was terminated by adding F12K with 10% FBS, followed by filtration with a 100 μm cell strainer (VWR). Pelleted crypts were resuspended in human 3D Organoid Culture Medium (HCM, Table S6) and Matrigel™ with a 1:5 volume ratio and embedded with 10–20 crypts per 10 μl droplet. Human organoids were expanded in HCM and differentiated in Human 3D Organoid Differentiation Medium (HDM, Table S6) for 72 hours.
Mouse and human colonic tissues used for experimentation were generally taken from proximal colon unless indicated otherwise.

Gu W., Wang H., Huang X., Kraiczy J., Singh P.N., Ng C., Dagdeviren S., Houghton S., Pellon-Cardenas O., Lan Y., Nie Y., Zhang J., Banerjee K.K., Onufer E.J., Warner B.W., Spence J., Scherl E., Rafii S., Lee R.T., Verzi M.P., Redmond D., Longman R., Helin K., Shivdasani R.A, & Zhou Q. (2021). SATB2 preserves colon stem cell identity and mediates ileum-colon conversion via enhancer remodeling. Cell stem cell, 29(1), 101-115.e10.

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Isolation of Murine Lung and Spleen Cells

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Lungs were removed aseptically, washed in R0L (RPMI-1640 (Sigma) + 2 mM L-Glutamine [Fisher Scientific, Loughborough, UK] + 50 U/mL penicillin [Sigma]) and cut into small pieces with sterile scissors. Lungs were incubated for 40 minutes at 37 °C (5% CO₂) with deoxyribonuclease I from bovine pancreas (type IV, Sigma; final concentration: 12 μg/mL) and Liberase TL (Sigma; final concentration: 625 μg/mL). The enzyme reaction was terminated by adding R10L (R0L + 10% heat-inactivated FBS [Sigma]) and homogenised lungs were passed through a 100-μm cell strainer (VWR International, Lutterworth, UK) to obtain a single cell suspension. Red blood cell lysis was performed using ammonium–chloride–potassium (ACK) lysis buffer. Spleens were removed aseptically and homogenised by passing through a 100-μm cell strainer. Red blood cell lysis was performed using RBC lysis buffer (Sigma). Processing of spleens was performed in R10S (antibiotic-free R10L). For all assays, samples from six mice were used to generate one pool per group.

Painter H., Prabowo S.A., Cia F., Stockdale L., Tanner R., Willcocks S., Reljic R., Fletcher H.A, & Zelmer A. (2020). Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells. Scientific Reports, 10, 3311.

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Isolation of Aortic Valve Interstitial Cells

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Porcine aortic heart valves from 4 animals were obtained from a local abattoir, transported to the lab on ice, and were digested to extract AVICs using previously published methods [39 (link)]. The AVs were washed in Earle’s Balanced Salt Solution (EBSS, Thermo Fisher Scientific, Waltham, MA) and then digested using a collagenase solution (0.75 mg/ml of collegenase in EBSS, Thermo Fisher Scientific) for 30 min. The initial digestion was intended to remove the valvular endothelial cells on the outside of the valves and this solution was discarded. Another round of digestion with fresh collagenase solution was performed for one hour before vortexing the solution to dislodge the AVICs. The AVICs were then separated using a 100 μm cell strainer (VWR, Radnor, PA) and the filtered solution was centrifuged. The resulting cell pellet was resuspended in standard growth media (10% FBS, 2% pen-strep, 0.4% fungizone, all Thermo Fisher Scientific). Only AVICs with passage numbers 2 – 4 were used to avoid senescence, genetic instability, and phenotype drift [40 (link)].

Khang A., Nguyen Q., Feng X., Howsmon D.P, & Sacks M.S. (2022). Three-dimensional analysis of hydrogel-imbedded aortic valve interstitial cell shape and its relation to contractile behavior. Acta biomaterialia, 163, 194-209.

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Embryonic Mouse Neuron Culture

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Cortices were dissected from E15 Swiss-Webster embryos in ice-cold HBSS (Thermo Fisher Scientific, 14175103) and dissociated with papain (Worthington Biochemical Corp, LS003126) and DNase I (Roche, 10104159001). Cells were resuspended in plating media (Neurobasal media (Thermo Fisher Scientific, 21103049), 1% Penicillin/Streptomycin Solution (Gemini Bio-Products, 400-109), 10% FBS) and filtered through a 100 μM cell strainer (VWR, 21008-950). Cell density was quantified using a Countess II Automated Cell Counter (Thermo Fisher Scientific, AMQAX1000), then plated on poly-D-Lysine-coated 12-well culture dishes at 0.5×106 cells or in 100mm culture dishes at 5×106 cells. Cultures were maintained in 5% CO2 at 37 °C in a cell culture incubator. After allowing four hours for the cells to adhere to the plate, the media was replaced and maintained with Neurobasal media supplemented with B-27 (Invitrogen, 17504-044), 1% Penicillin/Streptomycin, and 1% GlutaMAX Supplement (Thermo Fisher Scientific, 35050–079).

Cruz M.C., Sia R.A., Olson M., Cox G.M, & Heitman J. (2000). Comparison of the Roles of Calcineurin in Physiology and Virulence in Serotype D and Serotype A Strains of Cryptococcus neoformans. Infection and Immunity, 68(2), 982-985.

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Isolation of Porcine Aortic Valve Interstitial Cells

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Porcine aortic heart valves from 4 animals were obtained from a local abattoir, transported to the lab on ice, and were digested to extract AVICs using previously published methods [39] (link). The AVs were washed in Earle's Balanced Salt Solution (EBSS, Thermo Fisher Scientific, Waltham, MA) and then digested using a collagenase solution (0.75 mg/ml of collegenase in EBSS, Thermo Fisher Scientific) for 30 min. The initial digestion was intended to remove the valvular endothelial cells on the outside of the valves and this solution was discarded. Another round of digestion with fresh collagenase solution was performed for one hour before vortexing the solution to dislodge the AVICs. The AVICs were then separated using a 100 μm cell strainer (VWR, Radnor, PA) and the filtered solution was centrifuged. The resulting cell pellet was resuspended in standard growth media (10% FBS, 2% pen-strep, 0.4% fungizone, all Thermo Fisher Scientific). Only AVICs with passage numbers 2 -4 were used to avoid senescence, genetic instability, and phenotype drift [40] (link).

Khang A., Nguyen Q., Feng X., Howsmon D.P, & Sacks M.S. (2023). Three-dimensional analysis of hydrogel-imbedded aortic valve interstitial cell shape and its relation to contractile behavior. Acta biomaterialia, 163.

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