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Cellcrown

Manufactured by Merck Group
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Sourced in United Kingdom

The CellCrown is a laboratory equipment designed for cell culture applications. It provides a stable and controlled environment for the growth and maintenance of cells. The product's core function is to support cell culture processes by offering a reliable and consistent platform for cell cultivation.

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Lab products found in correlation

Trypan blue staining, by Thermo Fisher Scientific (1 mentions) Bepicm, by ScienCell (1 mentions) Pneumacult ali, by STEMCELL (1 mentions) Fitc dextran, by Merck Group (1 mentions) Ix50 s8f2, by Olympus (1 mentions) Penicillin streptomycin pen strep, by Thermo Fisher Scientific (1 mentions) Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions) Pen strep, by Merck Group (1 mentions) Polycarbonate filter, by Merck Group (1 mentions) Cellstar, by Merck Group (1 mentions)

Close spelling variants of this product

CellCrown™ inserts

7 protocols using cellcrown

1

Air-Liquid Interface Culture of Human Bronchial Epithelial Cells

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HBECs (Sciencell, California, USA) were cultured in bronchial epithelial cell medium (BEpiCM, Sciencell, California, USA), trypsinized and viable cells counted using trypan blue staining (Life Technologies, Paisley, UK). Cells were seeded to be confluent at 24 h onto either Falcon 0.4 μm pore size polyethylene terephthalate (PET; Becton Dickinson Labware, Claix, France) as a positive control or POSS-PCU sheets clipped with Cellcrown (Sigma–Aldrich, Gillingham, UK) to create a POSS-PCU transwell insert. 500 μL of Pneumacult-ALI (STEMCELL Technologies, Cambridge, UK) was added into the lower chamber of each well while cells were seeded in a 200 μL volume in the upper compartment. After three days of submerged culture, scaffolds were air-lifted by adding 1 mL of Pneumacult-ALI to the basal chamber and removing media from the upper chamber to generate an air-liquid interface.
Crowley C., Klanrit P., Butler C.R., Varanou A., Platé M., Hynds R.E., Chambers R.C., Seifalian A.M., Birchall M.A, & Janes S.M. (2016). Surface modification of a POSS-nanocomposite material to enhance cellular integration of a synthetic bioscaffold. Biomaterials, 83, 283-293.
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2

Quantifying Intestinal Permeability via Dextran

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Piglet scaffolds were mounted into CellCrowns (Sigma cat.no. Z742381) (instead of onto custom made mini-platforms) before seeding with organoids and culture as described above. Grafts in culture (or unseeded controls) were removed from culture and washed in PBS three times before 100μl of 500μg/ml FITC-Dextran (Sigma cat.no. 46944-100MG-F) in media was added to the apical chamber of the CellCrown. 900μl of media was added to the basal chamber. Samples were incubated for 2 hours at 37°C. Media was then collected from the basal chamber into individual wells of a flat-bottomed, black-walled 96-well plate in duplicate. Presence of FITC-Dextran was determined by measuring fluorescence (excitation 485nm, emission 535nm). Percentage leakage/permeability of FITC-Dextran through the grafts (from the “luminal side” to the “serosal side”) was calculated by comparison to empty well. Percentage leakage of blank scaffold was used as baseline leakage.
Laween M., Isobel M., Sara C., Weston A.E., Riana G., Lucinda T., Peter F., Michael O., Anna K., Anna B., Laura N., Nikolaos A., Elizabeth H., Julia K., Tedeschi A.M., Pellegata A.F., Susanna E., Alessandro A.P., Lucy C., Nikhil T., Thomas G.M., Simon E., Paola B., De Coppi P, & Li V.S. (2020). Engineering transplantable jejunal mucosal grafts using patient-derived organoids from children with intestinal failure. Nature medicine, 26(10), 1593-1601.
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3

Evaluating Electrospun Membrane Barrier Function

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The in vitro barrier function of the blending electrospun membranes was evaluated using L929 cells as a cellular model. The electrospun membranes of different groups were cut into circular disks (diameter, 25 mm) and then fixed in a Cell Crown™ (Sigma-Aldrich Co.). The fixed membranes were sterilized by immersing in 75% ethanol for 1 hour, rinsed three times with PBS and placed into a 24-well culture plate. A suspension of L929 cells in culture medium containing DMEM, 10% (v/v) FBS and 1% penicillin– streptomycin was seeded onto the membrane surfaces at a density of 3.0×104 cells/mL. After incubating at 37°C under a 5% CO2 atmosphere for 1 day, 4 days and 7 days, the fixed membranes were drawn out to observe the penetration results in the reverse side of membranes and the bottom of the culture plates by SEM and inverted phase contrast microscopy (Olympus IX50-S8F2), respectively.
Huang Y., Shi R., Gong M., Zhang J., Li W., Song Q., Wu C, & Tian W. (2018). Icariin-loaded electrospun PCL/gelatin sub-microfiber mat for preventing epidural adhesions after laminectomy. International Journal of Nanomedicine, 13, 4831-4844.
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4

Rat Mesenteric Tissue Culture Protocol

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All animal experiments were approved by Tulane University’s Institutional Animal Care and Use Committee. Rat mesenteric tissues were harvested and cultured according to our previous description [8 (link)]. Adult male Wistar rats (325–349 g) were anesthetized via intramuscular injection with ketamine (80 mg/kg body weight) and xylazine (8 mg/kg body weight). The mesentery was aseptically exteriorized [10 (link)] and rats were euthanized by intracardiac injection of 0.2 ml Beuthanasia. Then, mesenteric windows, defined as the thin, translucent connective tissues found between artery/vein pairs feeding the small intestine, were harvested, starting from the ileum. Tissues were immediately rinsed in sterile phosphate-buffered saline (PBS; Gibco; Grand Island, NY) with CaCl2 and MgCl2 at 37°C, and immersed in sterile minimum essential media (MEM; Gibco; Grand Island, NY) containing 1% Penicillin-Streptomycin (PenStrep; Gibco; Grand Island, NY). They were then transferred to individual wells in either a 6-well or 12-well culture plate. For drug studies, each tissue was quickly spread out on the bottom of a well and secured in place with a commercially available insert (CellCrown; Sigma-Aldrich; St. Louis, MO) with polycarbonate filter and covered with 3 ml of MEM containing 1% PenStrep. Tissues were cultured in standard incubator conditions (5% CO2).
Azimi M.S., Myers L., Lacey M., Stewart S.A., Shi Q., Katakam P.V., Mondal D, & Murfee W.L. (2015). An Ex Vivo Model for Anti-Angiogenic Drug Testing on Intact Microvascular Networks. PLoS ONE, 10(3), e0119227.
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5

Rat Mesenteric Tissue Preparation for Cell Printing

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All experiments were performed in accordance with the guidelines of the Tulane University Institutional Animal Care and Use Committee. Rat mesenteric tissue windows, defined as the thin connective tissue between the artery/vein pairs feeding the small intestine, were harvested from the ileum of adult male Wistar rats (Harlan, Indianapolis, IN) and incubated at 37°C/5% CO2/100% RH in MEM prior to LDW preparation. The mesentery tissues, spanning a triangular area of ~1.5 cm2 and a thickness of 20–40 μm, were then removed from media and stretched on inverted transwell inserts (CellCrown, Sigma Aldrich) with polycarbonate filters (Millipore, Billerica, MA). Surface tension interactions physically held tissues flat against the transwell filter membranes. The tissues were then rehydrated with 50 μL MEM and incubated in culture conditions for 20 mins. Excess media was removed prior to cell printing.
Phamduy T.B., Sweat R.S., Azimi M.S., Burow M.E., Murfee W.L, & Chrisey D.B. (2015). Printing Cancer Cells into Intact Microvascular Networks: A Model for Investigating Cancer Cell Dynamics during Angiogenesis. Integrative biology : quantitative biosciences from nano to macro, 7(9), 1068-1078.
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6

Peptide-SVP Co-Culture Scaffold Assay

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To test the cumulative effect of the peptides and the SVPs co-culture, control and Hep/Adh scaffolds were cut in 18 mm discs, locked in 24 well plate CellCrown™ (Sigma) inserts, and 1.5 x 105 Wheat Germ Agglutinin-AlexaFluor 568 (WGA-568) stained SVPs were seeded and let to attach for 2 hours on the Hep side. Then, 2.5 x 105 WGA-488 stained HUVECs were seeded on the Adh side by inverting the scaffold inside the CellCrowns™. Cells were cultured for 48 hours and then fixed and stained with DAPI. Number of nuclei was counted in three random fields acquired at 4x, using FIJI (ImageJ). Each experiment included at least two biological replicates and was repeated three independent times; each experiment was normalized to the control (control PCL without SVPs).
For gap closure experiment, a modification of the protocol above was applied wherein the 1.5 x 105 SVPs and 7.5 x 105 HUVECs were seeded on 6 well plate CellCrown™. Prior to HUVECs seeding a 0.5 mm PDMS barrier was locked in the middle of the PCL mat to prevent adhesion of the endothelial cells in the central region. After 3 hours, non-adherent HUVECs were washed away and the barrier was removed. Migration into the gap was assessed at 48 hours by fluorescence imaging of the whole well and quantified as mean grey value in 10 random fields of the gap area (n=3).
Campagnolo P., Gormley A.J., Chow L.W., Guex A.G., Parmar P.A., Puetzer J.L., Steele J.A., Breant A., Madeddu P, & Stevens M.M. (2016). Pericyte seeded dual peptide scaffold with improved endothelialization for vascular graft tissue engineering. Advanced healthcare materials, 5(23), 3046-3055.
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7

Antigen-Presenting Cell Culture on Scaffolds

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Scaffolds were cut in pieces of 6x6 mm per side, immobilized in 48 well-plates (CELLSTAR®) using CellCrown (Sigma Aldrich) and sterilized with a wash of Ethanol/Water 70% (v/v) and UV exposition of 15 min on each side. Three APC lines were seeded onto scaffolds at the concentration of 10,000 cells/well and tested as duplicates in the following in vitro assays.
Carrabba M., De Maria C., Oikawa A., Reni C., Rodriguez-Arabaolaza I., Spencer H., Slater S., Avolio E., Dang Z., Spinetti G., Madeddu P, & Vozzi G. (2016). Design, fabrication and perivascular implantation of bioactive scaffolds engineered with human adventitial progenitor cells for stimulation of arteriogenesis in peripheral ischemia. Biofabrication, 8(1).
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