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Cls3464

Manufactured by Corning
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The CLS3464 is a laboratory equipment product manufactured by Corning. It serves as a centrifuge for separating materials of different densities within a liquid sample. The core function of this product is to facilitate the separation and isolation of components within a mixture through the application of centrifugal force.

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Lab products found in correlation

Cell counting kit 8 (cck8), by Dojindo Laboratories (2 mentions) Paraformaldehyde (pfa), by Wuhan Servicebio Technology (2 mentions) Crystal violet staining solution, by Beyotime (2 mentions) Matrigel, by Corning (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) G1101, by Wuhan Servicebio Technology (1 mentions) Cell counting kit, by Yeasen (1 mentions) C0121, by Beyotime (1 mentions) Inverted light microscope, by Leica camera (1 mentions) C8470, by Solarbio (1 mentions) Bl539a, by Biosharp (1 mentions) L3755, by Merck Group (1 mentions) Dra00b, by R&D Systems (1 mentions) Tib 202tm, by American Type Culture Collection (1 mentions) F2442, by Merck Group (1 mentions)

12 protocols using cls3464

1

Evaluating Colorectal Cancer Cell Viability

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Cell viability was determined using MTT assay. Briefly, 1*103 colorectal cells were seeded into 96-well plates per well with 5 repetitions of each group. After 12 h of cell attachment, 10 µL Cell Counting Kit-8 (CK04, DOJINDO, Kumamoto, Japan) reagent (Cell Counting Kit, Yeasen) was added to 100 µL cell culture medium without FBS per well. Then, the plate was placed in an incubator at 37 °C for 2 h in the dark. Finally, the absorbance value of the whole plate was measured at 450 nm using a Biotek system. The curves of cell proliferation were plotted each day. As for colony-formation assays, 500 CRC cells were seeded into 6-well plates, and the colonies formed were counted 1 week later.
Transwell assays were performed by seeding 4*104–8*104 cells into the upper chamber (CLS3464, Corning Costar, Corning, NY, USA) with no FBS supplementation, while 500 µL DMEM with 10% FBS was added to the lower chamber. After 72 h of culture, migrated cells were fixed with 4% paraformaldehyde (G1101, Servicebio, Wuhan, Hubei, China), stained with crystal violet staining solution (C0121, Beyotime, Shanghai, China), and counted under a microscope.
Han L., Dai W., Luo W., Ye L., Fang H., Mo S., Li Q., Xu Y., Wang R, & Cai G. (2023). Enhanced De Novo Lipid Synthesis Mediated by FASN Induces Chemoresistance in Colorectal Cancer. Cancers, 15(3), 562.
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2

Gastric Cancer Cell Migration Assay

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In vitro experiments involved two human-derived gastric cancer cell lines: MKN-45 and SGC-7901. A control cell line (transfected with an empty vector) was established, along with two experimental cell lines (knockdown and overexpression groups). The knockdown group provided two stable cell lines constructed with shRNA sequences, while the overexpression group provided one stable cell line. The human GC cell line MKN-45 and SGC-7901 cell line were purchased from the National Cancer Institute (Bethesda, MD, USA). Transwell assays were performed by seeding 4 × 104–8 × 104 cells into the upper chamber (CLS3464, Corning Costar, Corning, NY, USA) with no FBS supplementation while the lower chamber was added 600 μL DMEM with 10% FBS. After 36–72 h of culture, migrated cells were fixed with 4% paraformaldehyde (G1101, Servicebio, Wuhan, Hubei, China), stained with Crystal Violet Staining Solution (C0121, Beyotime, Shanghai, China), and counted under a microscope. Transwell assays was repeated 3 times for each group, followed by statistical analysis. The statistical comparison was performed using a t-test, * indicating P value < 0.05; ** indicating P value < 0.01; *** indicating P value < 0.001.
Zhang H., Wang H., Ye L., Bao S., Zhang R., Che J., Luo W., Yu C, & Wang W. (2023). Comprehensive transcriptomic analyses identify KDM genes-related subtypes with different TME infiltrates in gastric cancer. BMC Cancer, 23, 454.
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3

Cell Viability and Migration Assay

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Cell Counting Kit‐8 (CK04, DOJINDO, Kumamoto, Japan) was used to evaluate cell viability. Transwell assays were performed by seeding 4 × 104‐8 × 104 cells into the upper chamber (CLS3464, Corning Costar, Corning, NY, USA) with no FBS supplementation while the lower chamber was added 500 μL DMEM with 10% FBS. After 36‐72 h of culture, migrated cells were fixed with 4% paraformaldehyde (G1101, Servicebio, Wuhan, Hubei, China), stained with Crystal Violet Staining Solution (C0121, Beyotime, Shanghai, China), and counted under a microscope.
Dai W., Xiang W., Han L., Yuan Z., Wang R., Ma Y., Yang Y., Cai S., Xu Y., Mo S., Li Q, & Cai G. (2022). PTPRO represses colorectal cancer tumorigenesis and progression by reprogramming fatty acid metabolism. Cancer Communications, 42(9), 848-867.
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4

Cell Migration Assay Protocols

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Cells were seeded into 24-well Corning transwell and incubated with control or MGH-CP1 at indicated concentrations (Corning Cat# CLS3464). Twenty-four hours later, migrating cells were stained with Crystal violet and quantitated.
The Oris Cell Migration Assay uses a 96-well plate with “stopper” barriers that create a central cell-free Detection Zone for cell migration experiments (PLATYPUS CAT# CMA1.101). Removing the stoppers allows the cells to migrate into the Detection Zone at the center of each well. Calcein-AM was applied to stain the viable cells.
Sun Y., Hu L., Tao Z., Jarugumilli G.K., Erb H., Singh A., Li Q., Cotton J.L., Greninger P., Egan R.K., Tony Ip Y., Benes C.H., Che J., Mao J, & Wu X. (2022). Pharmacological blockade of TEAD–YAP reveals its therapeutic limitation in cancer cells. Nature Communications, 13, 6744.
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5

Transwell Migration and Invasion Assay

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A transwell migration assay was performed using a 24 well plate and transwell inserts (Corning, CLS3464) containing microporous (8 μm pores) membranes. 40,000 cells were resuspended in serum free media, placed on top of each transwell, and allowed to migrate through the microporous (8 μm pore) membrane for 24 h towards the complete FBS-containing media. After 24 h, non-migratory cells on top of the membrane were removed using a cotton bud and membranes were stained with hematoxylin and eosin. Once dried, membranes were imaged at ×40 total magnification using the Leica Inverted Light Microscope and cells were counted using ImageJ software. For the invasion assay, transwells were coated with a thin layer of matrigel (BD Biosciences, 400 μg/ml diluted in serum free RPMI-1640 medium) for 4 h before performing the assay same as above for 48 h.
Halari C.D., Nandi P., Sidhu J., Sbirnac M., Zheng M, & Lala P.K. (2022). Decorin–induced, preeclampsia-associated microRNA-512-3p restrains extravillous trophoblast functions by targeting USF2/PPP3R1 axis. Frontiers in Cell and Developmental Biology, 10, 1014672.
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6

Cell Migration Assay Protocol

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Migration assays were performed using transparent polyethylene membrane cell culture inserts with an 8.0 µm pore size (Corning, CLS3464, USA) following the manufacturer’s instructions. HEI-OC1 cells (4 × 103) were seeded in 100 μl serum-free medium per well in the upper chamber, and medium containing 10% fetal bovine serum was added to the lower chamber (750 μl). After incubation at 33 °C for 24 h, the cells were fixed with 4% paraformaldehyde (Biosharp, BL539A, China) and stained with 0.1% crystal violet solution (Solarbio, C8470, China). Stained cells that migrated through the membrane were counted in five random areas under a microscope using a 20× objective. The cells at five randomly chosen areas were calculated for migration assays.
Liu R., Shang W., Liu Y., Xie Y., Luan J., Zhang T., Ma Y., Wang Z., Sun Y., Song X, & Han F. (2024). Inhibition of the ILK-AKT pathway by upregulation of PARVB contributes to the cochlear cell death in Fascin2 gene knockout mice. Cell Death Discovery, 10, 89.
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7

Cell Migration Assay for Mutant Cell Lines

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L6-WT, L6-V561M, H1581-WT, and H1581-V561M cells were serum starved for 12 h. 1×105 cells were then plated onto 8 μm-pore transwell migration filters in 24-well plates (Corning #CLS3464) in starvation media. Growth media (DMEM or RPMI-1640 with 10% FBS) was added to the bottom of the transwell and cells were incubated for either 8 h (L6 cells) or 24 h (H1581 cells) at 37°C under an atmosphere with 5% CO2. Unmigrated cells on the inner surface of the transwell chamber were removed, and migrated cells were stained with crystal violet. Assays were imaged using the Echo Revolve R4 microscope (San Diego, CA). Migrated cells were quantified using ImageJ and data were plotted using GraphPad Prism.
Ryan M.R., Sohl C.D., Luo B, & Anderson K.S. (2018). The FGFR1 V561M Gatekeeper Mutation Drives AZD4547 Resistance through STAT3 Activation and EMT. Molecular cancer research : MCR, 17(2), 532-543.
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8

THP-1 Macrophage Co-Culture Protocol

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THP-1 monocyte cells (tib-202tm; ATCC) were cultured in RPMI-1640 medium (12633012; Thermo Fischer) with 0.05 mM 2-mercaptoethanol and 10% fetal bovine serum (FBS). THP-1 cells were stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 hours to induce macrophage phenotype as assessed by adherence to tissue culture flask and expression of CD11b integrin (Figure S2). For co-culture, we followed an established protocol16 (link),22 (link). Briefly, ABCB5+ DSCs or donor-matched ABCB5− fractions were plated at 2x104 cells per well in 24-well plates in 0.5 ml Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS, 100 U/ml penicillin/streptomycin and 2 mM L-glutamine. After 24 hours, THP-1 derived macrophages were seeded on top or in transwell inserts (CLS3464; Corning) at 1×105 cells per well. Co-cultures were incubated with 50 U/ml recombinant human interferon-gamma (IFN-γ) (285-IF-100; R&D Systems) for 24 hours and then stimulated with 20 ng/ml LPS (L3755; Sigma-Aldrich) and 50 U/ml IFN-γ for another 24-hour period before supernatants were harvested and analyzed by enzyme linked immunosorbent assay (ELISA) for IL-1RA (DRA00B; R&D Systems). IL-1RA levels were analyzed between conditions with unpaired t-tests.
Riedl J., Pickett-Leonard M., Eide C., Kluth M.A., Ganss C., Frank N.Y., Frank M.H., Ebens C.L, & Tolar J. (2021). ABCB5+ dermal mesenchymal stromal cells with favorable skin homing and local immunomodulation for Recessive Dystrophic Epidermolysis Bullosa treatment. Stem cells (Dayton, Ohio), 39(7), 897-903.
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9

Boyden Chamber Assay for Cell Migration and Invasion

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Cell migration and invasion experiments were performed using the Boyden Chamber Assay (Corning, CLS3464). A total of 500,000 cells were seeded in serum-free RPMI media (Thermo Fisher Scientific, 118757093) in the upper chamber, and RPMI medium containing 10% FBS (Sigma-Aldrich, F2442) was placed in the lower chamber. For invasion experiments, the upper chamber was coated with Matrigel (Corning). After 4 hours, cells were treated with BAS-2 (30 μM). Following 24 hours (migration experiments) or 48 hours (invasion experiments) of treatment, cells were stained with 0.5% crystal violet.
Dowling C.M., Hollinshead K.E., Di Grande A., Pritchard J., Zhang H., Dillon E.T., Haley K., Papadopoulos E., Mehta A.K., Bleach R., Lindner A.U., Mooney B., Düssmann H., O’Connor D., Prehn J.H., Wynne K., Hemann M., Bradner J.E., Kimmelman A.C., Guerriero J.L., Cagney G., Wong K.K., Letai A.G, & Chonghaile T.N. (2021). Multiple screening approaches reveal HDAC6 as a novel regulator of glycolytic metabolism in triple-negative breast cancer. Science Advances, 7(3), eabc4897.
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10

Transwell Assay for Cell Migration

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Cell migration ability was evaluated by a 24-well transwell chamber with 8 µm pore size (Corning #CLS3464). Cells were suspended in 100 μl serum-free medium and added to the upper chamber, and cultured for 4 h. The lower chamber was then filled with 500 μl medium containing 10% FBS, and the upper chamber was inserted into a lower chamber, 8 h after incubation. Non-migrated cells were wiped from the upper surface of the transwell membrane, and cells that migrated to the underside of the membrane were fixed with methanol and stained with 20% Giemsa and counted under a microscope.
Chen J., Zeng Y., Wu R., Xuan Y., Jiang M, & Teng H. (2021). Decreased DUSP26 Expression Promotes Malignant Behavior in Glioblastoma Cells via Deregulation of MAPK and Akt Signaling Pathway. Frontiers in Oncology, 11, 622826.
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