Cell strainer

Manufactured by BD

Sourced in United States, United Kingdom, Germany, Japan, Canada, Sweden, France, Australia, Belgium, Italy

The cell strainer is a laboratory device used to filter and separate cells from a cell suspension. It consists of a mesh screen or filter that allows smaller particles, such as cells, to pass through while retaining larger debris or cell aggregates. The core function of the cell strainer is to provide a simple and efficient way to purify and prepare cell samples for downstream applications.

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70 and 40 μm cell strainers (6 citations) 70 µm and 40 µm cell strainers (2 citations) Sterile 70nm cell strainers (2 citations) 75- and 40-μm cell strainers (2 citations) 70-μm and 40-μm cell strainers (2 citations) 40-micron nylon cell strainers (2 citations) 100mm cell strainers (2 citations) 100-μm and 40-μm cell strainers (2 citations) 70-μm-pore nylon cell strainers (2 citations) Cell strainers (100-μm) (2 citations)

717 protocols using cell strainer

Isolation and Culture of ADSCs from Adipose Tissue

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The ADSCs from adipose tissue were isolated as previously described [19 (link)]. Briefly, the fat tissues were minced with a sterile blade, washed with phosphate buffered saline (PBS) three times, and digested with 0.1% collagenase type 1 (Invitrogen, Carlsbad, CA, USA) in a shaking incubator at 250 rpm and 37 °C for 60 min. The digested tissues were then centrifuged at 1500 rpm for 10 min to remove the remaining adipose tissue and oil. The pellet was incubated with red cell lysis buffer (Roche) for 10 min and filtered using a cell strainer (100 μm, Falcon, Corning, NY, USA), followed by centrifugation at 2000 rpm for 5 min. The pellet was resuspended in PBS, filtered with a cell strainer (70 μm, Falcon), and centrifuged at 2000 rpm for 5 min. The supernatant was discarded, and the cell pellet was resuspended in low glucose Dulbecco’s modified Eagle medium (DMEM) medium containing 15% fetal bovine serum (FBS, Invitrogen, Thermo Fisher Scientific Inc., Waltham, MA, USA), 100 U/mL penicillin (Invitrogen), 100 μg/mL streptomycin (Invitrogen), and 2 mM L-glutamine (Invitrogen) and incubated at 37 °C and 5% CO₂. After 48 h, the medium was changed and ADSC (passage 0) were maintained for 6–8 days. Passage numbers 2–5 of the ADSC were used for all experiments.

Park J.H., Choi Y.J., Kang S.Y., Ju H., Min K.W., Kim N.Y., Park H.Y., Kim E.S., Kwon M.J, & Suh Y.J. (2022). Organ-Specific Differentiation of Human Adipose-Derived Stem Cells in Various Organs of Xenotransplanted Rats: A Pilot Study. Life, 12(8), 1116.

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Lung Immune Cell Isolation and Analysis

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Mice were euthanized by carbon dioxide inhalation (100% CO₂ at slow fill rate of 20% of the chamber volume/minute) and lungs were perfused with PBS prior to collection. The spleen and medLNs were also collected from animals. Lungs were minced and incubated with Collagenase 3 (Scimar CLS-3, Worthington) and DNase I (Roche) in serum-free RPMI at 37°C for 60 min, then passed through a cell strainer (FALCON). Single cell suspensions were also prepared from spleen and medLNs by mechanical disruption through a cell strainer (FALCON). Cell suspensions from lungs, spleen and medLNs were treated with Red Blood Cell Lysing buffer (Sigma Aldrich) to lyse erythrocytes and then cell number and viability were determined using a hemocytometer and tryphan blue exclusion. Cell suspensions were then centrifuged and cells resuspended in FACS buffer [PBS, 2 mM EDTA, 2% FCS (Gibco)] to assess expression of cell-surface markers or in c-RPMI [RPMI 1640 (Gibco), 1 mM sodium pyruvate (Gibco), 2 mM L-glutamine (Gibco), 100 U/mL penicillin, 100 μg/mL streptomycin (Media Unit at Peter Doherty Institute, Melbourne), 5% FCS] for peptide stimulation followed by intracellular cytokine staining, as described below.

Fuglsang E., Pizzolla A., Krych L., Nielsen D.S., Brooks A.G., Frøkiær H, & Reading P.C. (2018). Changes in Gut Microbiota Prior to Influenza A Virus Infection Do Not Affect Immune Responses in Pups or Juvenile Mice. Frontiers in Cellular and Infection Microbiology, 8, 319.

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Intracellular ROS Measurement in Islets

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To measure the intracellular ROS in isolated islets, the islets were incubated in serum-free RPMI with DHE (final concentration of 10 µM) for 30 min in a noncoated dish. The islets were dissociated into single cells with 0.25% trypsin-EDTA (HyClone) in a rotator in a 37°C incubator for 5 min. The dissociated islet cells were dispersed in phenol-free RPMI containing 0.1% BSA and filtered with a round-bottom tube with a cell strainer (Falcon), and the intensity of DHE fluorescence was measured with flow cytometry (BD LSRFortessa).
The ROS concentrations in hydrogen peroxide–treated Tph1-null MIN6 cells were measured by 2,7-dichlorofluorescein diacetate (DCF-DA) assay. Tph1-null MIN6 cells were treated with 5-HT, 5-HTP, or N-acetylcysteine (NAC) for 24 hours in serum-free conditions and then treated with DCF-DA for 30 min (final concentration, 50 µM). The media were discarded, and the cells were washed once in PBS. The cells were incubated in DMEM with or without hydrogen peroxide (200 µM) containing 5-HT (500 µM), 5-HTP (200 µM), or NAC (5 mM). After 30 min of incubation, the cells were detached with trypsin-EDTA. The cells were filtered with a round-bottom tube with a cell strainer (Falcon), and the intensity of DCF-DA fluorescence was measured by flow cytometry (BD LSRFortessa).

Moon J.H., Kim H., Kim H., Park J., Choi W., Choi W., Hong H.J., Ro H.J., Jun S., Choi S.H., Banerjee R.R., Shong M., Cho N.H., Kim S.K., German M.S., Jang H.C, & Kim H. (2020). Lactation improves pancreatic β cell mass and function through serotonin production. Science translational medicine, 12(541), eaay0455.

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Isolation of Epidermal and Dermal Cell Suspensions

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Epidermal cell suspensions were prepared as previously described with slight modifications (Nagao et al., 2009 (link)). The shaved mouse trunk skins were floated on RPMI 1640 medium containing 0.15% trypsin and 0.27 mM EDTA at 37 °C for 30 or 45 min. The epidermis was mechanically scraped off in 5% FCS-PBS and washed and filtered through cell strainers (BD Falcon). To obtain the dermal cell suspension, the dermis was minced with scissors or scalpels and incubated in RPMI 1640 medium containing 0.25 mg/ml of Liberase TL Research grade (Roche) and 200 U/ml of DNase I (Sigma) at 37 °C for 60 min. Skin-draining lymph nodes (brachial, axiliary, cervical and inguinal) and spleens were excised and disrupted through a cell strainer (BD Falcon) in 5% FCS-PBS to generate single cell suspensions.

Kitashima D.Y., Kobayashi T., Woodring T., Idouchi K., Doebel T., Voisin B., Adachi T., Ouchi T., Takahashi H., Nishifuji K., Kaplan D.H., Clausen B.E., Amagai M, & Nagao K. (2017). Langerhans Cells Prevent Autoimmunity via Expansion of Keratinocyte Antigen-Specific Regulatory T Cells. EBioMedicine, 27, 293-303.

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Isolation of Bone-Derived Cells

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Femurs, tibias and ilium were dissected and crushed with a scissors and a pestle. The crushed bones were gently washed once in HBSS+ (Hanks-balanced salt solution supplemented with 2% FBS, 10 mM Hepes, and 100 U/mL penicillin/ 0.1 mg/mL streptomycin solution), and the solution filtered through a cell strainer (BD Falcon) was discarded. The bone fragments were collected and incubated for 1 h at 37 °C in 20 mL of DMEM (Invitrogen) containing 0.2% collagenase (Wako Chemicals USA, Inc.), 10 mM Hepes and 100 U/mL penicillin/ 0.1 mg/mL streptomycin solution. The suspension was filtered with a cell strainer (BD Falcon) to remove debris and bone fragments, and collected by centrifugation at 400 g for 5 min at 4 °C. The pellet was immersed in 1 mL water (Sigma-Aldrich) for 5–10 s to burst the red blood cells, after which 1 mL of 2 × PBS (diluted the product from Sigma-Aldrich) containing 4% FBS was added, and the suspension was filtered through a cell strainer. These serial procedures are described in previous report^{28 (link)}.

Kanazawa S., Okada H., Hojo H., Ohba S., Iwata J., Komura M., Hikita A, & Hoshi K. (2021). Mesenchymal stromal cells in the bone marrow niche consist of multi-populations with distinct transcriptional and epigenetic properties. Scientific Reports, 11, 15811.

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Isolation of Hepatocytes and Splenocytes

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Livers were extracted and perfused with collagenase type I (1mg/ml in RPMI) via portal vein injection until visible blanching of all lobes was observed. Liver lobes were then finely cut into small pieces using a razor blade and incubated for 10 minutes at 37°C with CO₂. They were then mashed through a cell strainer (Falcon, 70 μm) into a 50 ml conical and washed with ~40 ml of plain RPMI. Cells were spun in a tabletop centrifuge (Eppendorf, model 5810R) at 50 × g for 5 minutes at 4°C. The supernatant was discarded, the pellet resuspended in 15 ml of RPMI, and spun for an additional 2× at 50 × g for 5 minutes at 4°C. The cell pellet, now consisting of >96% hepatocytes (as previously reported⁵¹ and validated by flow cytometry) were resuspended once in PBS and spun again as before. Cells were resuspended in RBC Lysis Buffer, incubated for 5 minutes at room temperature (~22°C), spun at 1500 rpm in a small tabletop centrifuge (Eppendorf model 5810R), and the pellet resuspended in RPMI or 1X PBS depending on usage. For splenocyte isolation: spleens were mashed through a cell strainer (Falcon, 70 μm) into a 50 ml conical and washed with 15 ml of plain RPMI. Cells were spun at 1,000 × g for 5 minutes, once in RPMI, and once in PBS. They were then RBC lysis treated, spun, and the pellet resuspended in RPMI or 1 X PBS.

Nozaki K., Maltez V.I., Rayamajhi M., Tubbs A.L., Mitchell J.E., Lacey C.A., Harvest C.K., Li L., Nash W.T., Larson H.N., McGlaughon B.D., Moorman N.J., Brown M.G., Whitmire J.K, & Miao E.A. (2022). Caspase-7 activates ASM to repair gasdermin and perforin pores. Nature, 606(7916), 960-967.

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DNA Extraction from Leaf Samples

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DNA was extracted from 3.5 g of fresh intact leaves, frozen, then crushed in liquid nitrogen. Cell nuclei were first purified. The powdered tissue was suspended in 45 mL of Sucrose Extraction Buffer (SEB) containing 0.01 M Tris, 0.1 M KCl, 0.01 M EDTA (0.5 M, pH 8), 0.55 M sucrose, 5 mM spermidine, 0.13% carbamic acid, 0.25% PVP 40, and 0.2% β-mercaptoethanol, before incubation on ice for 12 min. The supernatant was filtered through Miracloth (475,855, Merck Millipore®, France) then through a cell strainer (40 μm, Falcon®, Germany) and incubated on ice for 12 min before adding 0.15 vol. of SEB with 10% triton (X 100). Incubation on ice was continued for 12 min before centrifugation (600 g, 10 min, 4 °C). The pellet was resuspended in 60 mL of SEB for an additional filtration on a cell strainer (40 μm, Falcon®, Germany) before incubation on ice and centrifugation in the same conditions. The pellet was resuspended in 2 mL of SEB. Nuclei were lysed by adding 3 vol. of TBL buffer containing 0.4 M EDTA (0.5 M, pH 8), 2% N lauryl sarkosyl and 1 mg.mL⁻¹ Proteinase K, before incubation at 55 °C for 3 h with gentle shaking. DNA extraction and purification were performed using the DNeasy™ Plant Maxi kit (Qiagen, France) following the manufacturer’s recommendations. DNA extracts were stored at −20 °C.

Maillot P., Velt A., Rustenholz C., Butterlin G., Merdinoglu D, & Duchêne E. (2021). Alternative splicing regulation appears to play a crucial role in grape berry development and is also potentially involved in adaptation responses to the environment. BMC Plant Biology, 21, 487.

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Isolation and Characterization of Immune Cells

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Blood samples were anti-coagulated with Na₂EDTA and resuspended in 0.84% NH₄Cl solution to lyse the red blood cells (RBCs). Peripheral blood mononuclear cells (PBMCs) were collected after washing twice with PBS and the number of cells was calculated using cell count plates.
Lung lymphocytes were isolated as previously described^{70 (link)} Mice were exsanguinated from the orbital cavity to minimize the amount of blood in the lungs. To obtain a single-cell suspension, the lungs were removed and passed through a cell strainer (BD Falcon). The cells were resuspended in 35% Percoll solution (in PBS buffer) and centrifuged at 1,500 rpm for 15 min at room temperature. Red blood cells in the lymphocyte pellet were lysed with 0.8% NH₄Cl and washed twice with PBS.
Spleens were removed and passed through a cell strainer (BD Falcon) to obtain a single cell suspension, and the spleen leukocytes were isolated with 0.84% NH₄Cl to lyse erythrocytes.

Hu X., Sun X., Zhao Y., Iv C., Sun X., Jin M, & Zhang Q. (2023). GlcNac produced by the gut microbiome enhances host influenza resistance by modulating NK cells. Gut Microbes, 15(2), 2271620.

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Splenocyte Isolation and RBC Lysis

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Spleens were smashed using a 70 µm Cell Strainer (Falcon, Becton Dickinson) and the splenocytes were collected using cold RPMI-1640 medium (Gibco, Life Technologies). Splenocytes were centrifuged at 300 g for 10 min at room temperature and the pelleted cells were treated with a red blood cell lysis buffer (BioLegend), according to the manufacturer’s instructions. Obtained splenocytes were filtered with a 70 µm Cell Strainer (Falcon, Becton Dickinson).

Vo H.T., Baudner B.C., Sammicheli S., Iannacone M., D’Oro U, & Piccioli D. (2018). Alum/Toll-Like Receptor 7 Adjuvant Enhances the Expansion of Memory B Cell Compartment Within the Draining Lymph Node. Frontiers in Immunology, 9, 641.

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Isolation of Immune Cells from Murine Tissues

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Bone marrow cells were flushed thoroughly from mouse femurs with 10 ml of PBS. Peripheral blood leukocytes were obtained through retro-orbital bleeding by heparinized capillary tubes. Single cell suspensions from spleens were prepared by mashing through 70 μM Cell Strainers (BD) without enzymatic digestion. Lungs were perfused via the right ventricle of the heart with PBS. The tissues were minced, digested in 0.4mg/ml collagenase IV (Sigma) for 45 minutes and filtered through 70 μM Cell Strainer (BD) to get the cells suspension. Red blood cells from the bone marrow, peripheral blood, spleen and lungs were removed by RBC lysis buffer (eBioscience). For liver leukocyte isolation, livers were perfused in situ via the portal vein with PBS. Perfused livers were dissected and digested with 0.1mg/ml collagenase IV (Sigma), except in cases where annexin V staining was to be performed. The resultant cell suspensions were layered onto a 2-step (40%/70%) discontinuous Percoll gradient (GE Healthcare) and centrifuged at 900g for 20 minutes at 25°C. Hepatic leukocyte populations collected at the interface were washed twice in wash medium and used for analysis.

Jiao J., Dragomir A.C., Kocabayoglu P., Rahman A.H., Chow A., Hashimoto D., Leboeuf M., Kraus T., Moran T., Carrasco-Avino G., Friedman S.L., Merad M, & Aloman C. (2014). Central Role of Conventional Dendritic Cells in Regulation of Bone Marrow Release and Survival of Neutrophils. Journal of immunology (Baltimore, Md. : 1950), 192(7), 3374-3382.

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