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Nylon mesh cell strainer

Manufactured by Corning
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The Nylon mesh cell strainer is a laboratory equipment designed for the filtration and separation of cells from a suspension. It features a fine nylon mesh that allows the passage of single cells while retaining larger cell aggregates or debris. This product is used to prepare single-cell suspensions for various cell-based applications, such as cell culture, flow cytometry, and cell sorting.

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Lab products found in correlation

Percoll solution, by Merck Group (1 mentions) Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions) Collagenase 4, by Merck Group (1 mentions) Fluorescence microscope, by Leica (1 mentions) Tumor dissociation kit, by Miltenyi Biotec (1 mentions) Cd45 microbeads, by Miltenyi Biotec (1 mentions) Dmem f12, by Thermo Fisher Scientific (1 mentions) Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (1 mentions) Collagenase 4, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Hyaluronidase, by Merck Group (1 mentions) Dnase 1, by Merck Group (1 mentions) Facsdiva software, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Rnaiso plus, by Takara Bio (1 mentions) Rnalater, by Merck Group (1 mentions) Nucleospin rna blood kit, by Macherey-Nagel (1 mentions)

Spelling variants of this product

Cell strainer (160 citations) Nylon cell strainer (42 citations) Nylon mesh (5 citations) Nylon mesh cell-strainer cap (3 citations) 70 μm nylon mesh cell strainer (3 citations) 35-μm nylon mesh cell strainer (2 citations)

9 protocols using nylon mesh cell strainer

1

Isolation and Culture of Primary Hepatocytes

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Livers were perfused in situ via the portal vein with Hanks balanced salt solution (HBSS; Gibco) supplemented with 5% heat‐inactivated FBS, followed by 0.3% collagenase IV (Sigma‐Aldrich). Perfused livers were dissected and teased through 70 µm nylon mesh cell strainers (Corning). Liver cells were suspended and centrifuged at 50 g for 2 min for 3 times.

The supernatant was collected by centrifugation at 800 g for 5 min. Thereafter, cells were suspended and allowed to attach to cell culture plates for 15 min at 37 C, and the attached cells were KCs.

Primary hepatocytes were pelleted after centrifugation at 50 g for 2 min. Cells were resuspended in 20 ml of 40% cold Percoll solution (Sigma‐Aldrich) and centrifuged at 150 g for 7 min. The pelleted hepatocytes were suspended in plating medium (Williams E medium with hepatocyte thawing and plating supplement pack; Gibco) and plated in collagen type I‐coated plates for 3 hr. Maintenance medium (Williams E medium with hepatocyte maintenance supplement pack; Gibco) was used for cultures overnight or longer.

Hepatocytes culture HR patterns were imposed following a method described previously (Strey et al., 2010).
Zhong W., Rao Z., Rao J., Han G., Wang P., Jiang T., Pan X., Zhou S., Zhou H, & Wang X. (2020). Aging aggravated liver ischemia and reperfusion injury by promoting STING‐mediated NLRP3 activation in macrophages. Aging Cell, 19(8), e13186.
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2

Isolation and Analysis of Liver Tumor Cells

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The isolated liver tumor was minced and incubated with the digestion solution (Tumor Dissociation Kit; Miltenyi Biotec, Bergisch Gladbach, Germany) at 37 °C for 1 h. A single cell suspension was generated by successively passing the digestion product through 70 µm and 40 µm nylon mesh cell strainers (Corning). Then, the ACK lysis buffer was applied to exclude red blood cells, and CD45 microbeads (Miltenyi Biotec) were used to remove leukocytes. Single cells were pipetted into individual polymerase chain reaction (PCR) tubes and analyzed under a fluorescence microscope (Leica, Wetzlar, Germany).
Chen L., Yi X., Guo P., Guo H., Chen Z., Hou C., Qi L., Wang Y., Li C., Liu P., Liu Y., Xu Y, & Zhang N. (2020). The role of bone marrow-derived cells in the origin of liver cancer revealed by single-cell sequencing. Cancer Biology & Medicine, 17(1), 142-153.
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3

Isolation and Culture of Mixed Glia

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Mixed glia cultures were prepared from WT C57BL/6 J and TRPV1-KO mouse pups as previously described [13 (link), 23 (link)]. Briefly, 1- to 2-day-old mouse pups were killed by decapitation, and their brains were dissected. The meninges were removed, and cerebral cortexes were cut into small pieces. The tissue pieces were homogenized and then filtered by passing through a nylon-mesh cell strainer (70 μM pores, Corning, United States). DMEM/F12 (Gibco, United States) supplemented with 10% FBS was added, and the cells were resuspended, and further were seeded into T75 flasks. Cells were then incubated in a thermostatic (37˚C) humidified CO2 (5%) incubator. The medium was changed every 3–4 days. About 12–15 days later, the matured microglia were separated by shaking off (200 rpm) for 2 h in a constant temperature shaker (37˚C), then the cells were collected, counted and seeded in the plates.
Zhang Y., Hou B., Liang P., Lu X., Wu Y., Zhang X., Fan Y., Liu Y., Chen T., Liu W., Peng B., Yin J., Han S, & He X. (2021). TRPV1 channel mediates NLRP3 inflammasome-dependent neuroinflammation in microglia. Cell Death & Disease, 12(12), 1159.
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4

Flow Cytometry Analysis of hPGCLC-EBs

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Day 6 hPGCLC-EBs were harvested and washed with DPBS (Invitrogen), then digested with an enzyme mix containing 5 mg/mL Collagenase IV (Invitrogen), 2.5 mg/mL Hyaluronidase (Sigma-Aldrich) and 100 U/mL Dnase I (Sigma-Aldrich) for 30 min at 37 °C. A single cell suspension of the digested EBs and of dissociated hiPSC F30 was created by pipetting up and down vigorously and passing through a 40 μm nylon mesh cell strainer (Corning). After washing with FACS buffer (DPBS (Invitrogen) with 0.5% bovine serum albumin (BSA) (Sigma-Aldrich)), cells were treated with conjugated antibodies (Table S1) for 30 min at 4 °C, washed with FACS buffer and resuspend in 200 µL FACS buffer with 1 μL 7AAD live/dead exclusion dye (BD Biosciences, San Jose, CA, USA). The EB cell suspensions were analyzed on an LSR-II flow cytometer (BD Biosciences). The FACS data were collected from FACSDiva Software (BD Biosciences) and analyzed with FlowJo software (BD Biosciences).
Chang Y.W., Overeem A.W., Roelse C.M., Fan X., Freund C, & Chuva de Sousa Lopes S.M. (2021). Tissue of Origin, but Not XCI State, Influences Germ Cell Differentiation from Human Pluripotent Stem Cells. Cells, 10(9), 2400.
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5

Dissociation and Culturing of HepaRG Cells

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Fully differentiated HepaRG cells were dissociated from tissue culture dishes using TrypLE™ Express. Following centrifugation at 300 × g for 5 min, the cells were resuspended in fresh differentiation medium and passed through a 40-μm nylon mesh cell strainer (Corning Inc., Corning, NY, USA) to minimize cell aggregation. Then cells at varying densities (5K (5,000), Umemura et al., 2007; (link)WHO, 2021 Monarca et al., 1991; (link)Goncharova et al., 1988; (link)Shelby et al., 1993; (link)NTP, 2022a (link)Nohmi, 2018; (link)Morita et al., 2016; (link)Ding et al., 2015; (link)Martins et al., 2012; (link)Schulte-Hubbert et al., 2020; (link)Muller et al., 1994; (link)Villarini et al., 2014 (link)Nohmi, 2018; (link)Mittelstaedt et al., 2004; (link)Culp, 2004 a Groups were classified by International Agency for Research on Cancer (IARC) Monographs on the identification of carcinogenic hazards to humans, Volumes 1-125 (https://monographs.iarc.who.int/list-of-classifications). +/-, both positive and negative results were reported. (Seo et al., 2020) (link).
Seo J.E., He X., Muskhelishvili L., Malhi P., Mei N., Manjanatha M., Bryant M., Zhou T., Robison T, & Guo X. (2022). Evaluation of an in vitro three-dimensional HepaRG spheroid model for genotoxicity testing using the high-throughput CometChip platform. ALTEX, 39(4).
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6

Single-cell isolation from murine spleen

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Single-cell suspensions were prepared in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Waltham, MA, USA or FUJIFILM Wako Chemicals, Osaka, Japan) supplemented with 1% heat-inactivated fetal calf serum (FCS) and 1 mM EDTA. To isolate mononuclear cells from the spleen, tissues were mashed using frosted glass microscope slides. Single cell suspension from the spleen was resuspended in Gey’s solution (130 mM NH4Cl, 5.0 mM KCl, 0.8 mM Na2HPO4, 0.18 mM KH2PO4, 5.6 mM Glucose, 0.03 mM Phenol red, 1.0 mM MgCl2, 0.3 mM MgSO4, 1.5 mM CaCl2, 13 mM NaHCO3) for red blood cell lysis. Cell debris was removed using a 40–100 µm nylon mesh cell strainer (CORNING, NY, USA).
Tamura Y., Yamane K., Kawano Y., Bullinger L., Wirtz T., Weber T., Sander S., Ohki S., Kitajima Y., Okada S., Rajewsky K, & Yasuda T. (2022). Concomitant Cytotoxic Effector Differentiation of CD4+ and CD8+ T Cells in Response to EBV-Infected B Cells. Cancers, 14(17), 4118.
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7

Isolation of Epididymal Stromal Vascular Fraction

Cited 2 times
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Epididymal stromal vascular fraction was isolated as previously described2 (link),18 (link). Briefly, AT was minced and digested with Collagenase type I (#LS004197, Worthington Biochemical Company, Lakewood, NJ, USA) (1 mg/ml) in KRH buffer (130 mM NaCl, 4.7 mM KCl, 1.24 mM MgSO4, 2.5 mM CaCl2, 1 mM HEPES, 2.5 mM NaH2PO4, 5 mM D-glucose, 200 nM adenosine, pH 7.4) containing 2.5% BSA for 45 min in a shaking incubator at 37°C. Cell suspension was then centrifuged at 400 × g for 5 min, treated with AKC lysis buffer (0.15 M NH4Cl, 0.01 M KHCO3, 0.1 mM EDTA) and filtered through a 70 μm nylon mesh cell strainer (Corning, NY, USA). Lysis was stopped using FACS buffer (0.1% NaN3, 1% BSA in PBS) and cells were centrifuged again.
Misiou A., Garmey J.C., Hensien J.M., Harmon D.B., Osinski V., McSkimming C., Marshall M.A., Fischer J.W., Grandoch M, & McNamara C.A. (2020). Helix-Loop-Helix Factor Id3: A Novel Regulator of Hyaluronan-mediated Adipose Tissue Inflammation. Arteriosclerosis, thrombosis, and vascular biology, 41(2), 796-807.
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8

Bone Marrow Transplantation in Mice

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Six-week-old GFP transgenic mice (male) were used as the bone marrow donor mice. Total bone marrow was flushed from the marrow cavity of the femurs and tibias using a 1 mL syringe, and successively passed through a 70 µm and 40 µm nylon mesh cell strainer (Corning, Corning, NY, USA) to produce a single cell suspension in phosphate-buffered saline (PBS). Then, the red blood cells were excluded using an ammonium chloride-potassium (ACK) lysis buffer. Recipient wild-type C57BL/6 mice (male) were irradiated twice with a dose of 4.5 Gys per time, from an X-ray irradiator. The interval was 4 h. A total of 1 × 106 (link) donor marrow cells were then injected once into the tail vein of these mice. After 4 weeks of recovery, these mice were used for further experiments.
Chen L., Yi X., Guo P., Guo H., Chen Z., Hou C., Qi L., Wang Y., Li C., Liu P., Liu Y., Xu Y, & Zhang N. (2020). The role of bone marrow-derived cells in the origin of liver cancer revealed by single-cell sequencing. Cancer Biology & Medicine, 17(1), 142-153.
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9

Isolating Immune Cells for LPS Response

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In both rat and mouse experiments, blood samples were collected under anesthesia 2 h after LPS administration, followed by euthanization and spleen isolation.
In the rat experiment, blood samples were collected in heparinized syringes, and leukocytes were isolated using erythrocyte lysis buffer, as described by Hoffman et al. [11] (link). The spleen was removed, and splenocytes were harvested by passing the cells through a nylon mesh cell strainer (Falcon-Corning, Corning, NY, USA). Splenocytes were further purified using Hoffman lysis buffer. The obtained splenocytes were used for the NK cell activity test and RT-qPCR. Splenocytes for RT-qPCR were washed with phosphate-buffered saline, lysed, and homogenized in RNAiso Plus (Takara Bio Inc., Kusatsu, Japan). A portion of the spleen was sliced and treated with RNAlater (Sigma-Aldrich, St. Louis, MO, USA) immediately after tissue removal. Treated tissue samples were stored at -70˚C until RNA extraction.
In the mouse experiment, blood samples were collected in heparinized syringes, treated with Buffer DL from the NucleoSpin RNA Blood Kit (Macherey-Nagel GmbH & Co. KG, Düren, Germany), and kept frozen at -70˚C until RNA extraction. The removed spleen was treated with RNAlater and processed as described above.
Mitsunaga F., & Nakamura S. (2021). A Sensitive and Simple Method to Assess NK Cell Activity by RT-qPCR for Granzyme B Using Spleen and Blood. Journal of Biosciences and Medicines, 09(03), 27-38.
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