Primary LUADs were pushed through 70 μm strainers (Corning) and cultured for 48 h in 96-U bottom plates in RPMI−1640 (Gibco) supplemented with 10% human serum, 1% L-glutamine, 1% penicillin and streptomycin, in the presence of control or siWnt1-loaded DOPC nanoliposomes (5ug per 4 × 106 cells).
70 μm strainer
Manufactured by Corning
Sourced in United States
The 70-μm strainer is a lab equipment product designed to filter and separate particles or cellular components from a liquid sample. It features a stainless steel mesh screen with a pore size of 70 micrometers, which allows the passage of smaller particles while retaining larger ones. This product is intended for use in various laboratory applications requiring filtration and sample preparation.
Automatically generated - may contain errors
Lab products found in correlation
Collagenase d, by Roche (2 mentions)
Dnase, by Roche (2 mentions)
Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (2 mentions)
Dispase, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Trypan blue, by Thermo Fisher Scientific (1 mentions)
Rbc lysis buffer, by BioLegend (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Matrigel, by Thermo Fisher Scientific (1 mentions)
Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions)
Dispase, by Merck Group (1 mentions)
Matrigel, by BD (1 mentions)
Dnase 1, by Roche (1 mentions)
Poly l lysine (pll), by Merck Group (1 mentions)
Trypsin, by Worthington (1 mentions)
Dnase 1, by Worthington (1 mentions)
Dnase type 1, by Merck Group (1 mentions)
Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (1 mentions)
Collagenase type 4, by Worthington (1 mentions)
17 protocols using 70 μm strainer
1
Lung Cancer Cells Treated with Wnt1 siRNA
2
Enzymatic Tissue Dissociation for Single-Cell Analysis
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Fresh tissue was transported in cold PBS on ice. Tissue was washed three times with PBS to remove blood, then cut into tiny pieces (∼1–2 mm3) and enzymatically dissociated by 1 mg/ml Type IV collagenase for 45 min at 37°C. The dissociation process was carried out in a shaking incubator with. Undigested tissue masses were removed using 70-μM strainers (Corning), and red blood cells (RBC) were removed by use of RBC lysis buffer (Biolegend). The dissociated cells were washed twice with PBS containing 0.04% bovine serum albumin (BSA, Sigma-Aldrich). Finally, cell viability was examined by Trypan blue (Invitrogen) staining.
3
Matrigel Plug Assay for Immune Cell Function
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PBMCs from healthy donors were stained and sorted as described previously. Blood cDC2 (105) and monocytes (3 × 105) were flow cytometry-sorted and injected subcutaneously along with B16_huGM (105) in 200 μL of ice-cold Matrigel® (BD Biosciences). Mice were sacrificed at day 4 by cervical dislocation and Matrigel® plugs were collected. Subcutaneous Matrigel® plugs were recovered, cut in pieces and incubated in HBSS (Life Technologies) 1% FBS, 0.37 U/ml Collagenase D (Roche), 10 μg/ml DNaseI (Roche) and 1 mg/ml Dispase (Sigma-Aldrich) for 30 minutes at 37°C. After digestion, plugs were smashed on a 70 μm strainer (Corning) and cells were collected and resuspended in flow cytometry buffer for flow cytometry analysis.
4
Culturing Primary Mouse Hippocampal Neurons
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Hippocampi from E18 mouse embryos were dissected free of meninges, minced, washed in HBSS and digested with 0.25% trypsin (Worthington) plus 0.05 mg/ml DNase I (Worthington) for 15 minutes at 37° C. trypsinization was stopped with DMEM supplemented with 10% FBS and tissue was washed twice and subsequently dissociated by triturating 10 times with a flamed Pasteur pipet. Cells were passed through a 70 μm strainer (Corning) and plated at a density of 7 × 105 cells/well in poly-L-lysine (Sigma) coated 6-well plates in DMEM supplemented with 10% FBS. Four hours later, medium was removed and replaced with Neurobasal medium supplemented with 2% B27 (Invitrogen) and 100 units/ml penicillin and 100 μg/ml streptomycin. Subsequently, half volume of Neurobasal medium in each well was replaced twice a week. After one week, hippocampal neurons were treated with 50 ng/ml recombinant soluble type I or type III for 48 hours, respectively. For all treatments, cells were treated in triplicates and experiments were repeated a minimum of three times to ensure reproducibility.
5
Isolation of Murine Immune Cells
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Mice were sacrificed by CO2 asphyxiation or injection of overdose (400 mg/kg of body weight) sodium pentobarbital (i.p. or i.v.). PEL was collected with 5 mM EDTA in DPBS (Thermo Fisher). AMs were isolated by washing the lungs three times with 0.4 mL of ice‐cold PBS containing 2 mM EDTA through an intratracheal cannula. All organs except the lung were removed prior to heart perfusion with DPBS. Lungs were digested for 45 min at 37°C in IMDM (Thermo Fisher) with 2 mg/mL of type IV collagenase (Worthington) and 0.02 mg/mL of type I DNase (Sigma). BM cells were flushed from tibias and femurs and passed through a 70‐μm strainer (Corning). All other organs were directly passed through 70‐μm strainer. Red blood cell lysis was performed with ACK buffer (0.15 M NH4Cl, 1 mM KHCO3, and 0.1 mM EDTA) on spleen and BM cells.
6
Isolation of Uterine Cells from Mice
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Female ICR mice (purchased from the Zhejiang Academy of Medical Science) of different ages (P0, P7, P14, P28, and P56) were utilized in this study. Uterus tissues were prepared as described previously (Masuda et al., 2007 (link)). Mice were euthanized by cervical dislocation, and whole uterus were explanted by microdissection and immersed immediately into ice-cold PBS (Sigma), followed by removal of the surrounding tissues such as blood vessels, ovaries, and oviducts. All procedures were performed with approved protocols in accordance with guidelines of the animal experimental center of Zhejiang University.
Single-cell suspensions of uterus tissues were then prepared by enzymatic digestion overnight at 4°C with type I collagenase (Life Technologies) diluted in low-glucose DMEM (Gibco), and the digested tissues were triturated into a single-cell suspension with a 1-mL pipette. The enzymes and cell debris were removed by centrifugation at 200 × g for 5 min and washed with PBS, followed by filtering the single-cell suspension through a 70-μm strainer (Corning). The resultant single-cell suspension could then be utilized for flow-cytometry analysis, single-cell analysis, and cell culture.
Single-cell suspensions of uterus tissues were then prepared by enzymatic digestion overnight at 4°C with type I collagenase (Life Technologies) diluted in low-glucose DMEM (Gibco), and the digested tissues were triturated into a single-cell suspension with a 1-mL pipette. The enzymes and cell debris were removed by centrifugation at 200 × g for 5 min and washed with PBS, followed by filtering the single-cell suspension through a 70-μm strainer (Corning). The resultant single-cell suspension could then be utilized for flow-cytometry analysis, single-cell analysis, and cell culture.
7
Isolation and Analysis of Tumor-Infiltrating Lymphocytes
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The tumors were harvested at day 21 and analyzed by flow cytometry. Tumor-infiltrating lymphocytes (TILs) were obtained using a previously described method (26 (link)). Briefly, the tumor was finely minced and incubated with 400(U/mL) Collagenase D (Roche) and (5 μg/mL) DNAse (Roche) for 1 hour at 37°C. Next, enzymatic digestion was stopped by adding 0.5μM EDTA, and digested tissues were filtered through a 70μm strainer (Corning). After that, the cell suspensions were treated with 0.2 mg/mL DNAse. The lymphocyte interface of the centrifuged Percoll 40/90 solution was recovered, washed, and stained as needed.
8
Tissue Dissociation and Single-Cell Preparation
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The lymph nodes and the ears were treated with 0.25 mg/mL liberase™ TL (Thermolysin low) (Roche) and 0.125 mg/mL DNAse (Roche) at 37°C for 25 minutes and 45 minutes, respectively. Tissues were chopped and incubated at the same conditions for 45 min more under constant shaking. Next, enzymatic digestion was stopped by adding 0.5μM EDTA, and cell suspensions were filtered through a 70 μm strainer (Corning). Afterwards, cells were washed with RPMI-1640 (Biowest) digestion medium containing 10% Fetal Bovine Serum (FBS) (Biowest), 2 mM L-glutamine (Corning), 100 IU Penicillin, and 100 μg/mL Streptomycin (Corning), by centrifugation at 400 g for 5 minutes. Then, 0.125 mg/mL DNAse was added, and cells were washed with digestion media for 5 minutes at 400 g. The supernatant was discarded, and cells were counted or stained, as needed.
9
Isolating Lung Cells from TB and HIV Patients
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Surgical samples from diseased lung tissue (n = 3 TB+HIV+; n = 3 TB+; n = 2 non-infected patients) were processed as described in (Ardain et al., 2019 (link)). Briefly, each sample was collected into cold RP-10 (RPMI (Sigma-Aldrich) + 10% FBS), minced, and incubated for 25-30 min at 37°C with digestion buffer containing collagenase D (Sigma-Aldrich), DNase I (Sigma-Aldrich) in RPMI 1640 (Sigma-Aldrich) with 10% FBS (Hyclone). Following incubation, samples were homogenized using a GentleMACS, filtered using a 70 μm metal strainer, and pelleted by centrifugation at 400 g for 5 min. After obtaining the pellet, cells were resuspended in RP-10, passed through another 70μm strainer (Corning), stained with trypan blue, and then counted and diluted for scRNA-seq using Seq-Well S3 (see below).
10
Isolation of Outer Root Sheath Cells
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Outer root sheath cells were isolated from the same hair specimens mentioned above. The hair follicle bulb was separated using microscissors and treated with 0.1% dispase (Invitrogen, CA, United States) for 45 min. The dermal sheaths were separated and removed under a stereoscope, and the epidermis was treated with 0.05% trypsin (Gibco) for 10 min. After terminating digestion, the samples were filtered through a 70-μm strainer (Corning Inc.) and cultured with defined keratinocyte-SFM (KSFM; Gibco) in flasks that were pre-coated with 10 μg/mL human fibronectin (Sigma-Aldrich, MO, United States).
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