Tissue culture was performed in a SterilGARD III Advance SG 603 laminar flow hood from Baker Company. Cell cultures were observed using a Zeiss Axiovert 25 inverted microscope. All cell lines and culture media were purchased from ATCC. Cell line HEK-293 (#CRL-1573) was grown in Eagle’s minimum essential medium (EMEM) (# 30–2003) and supplemented with 10% fetal bovine serum (FBS) and 1% Pen/Strep. Cell line MCF7 (ATCC HTB-22) was cultured in EMEM with 0.1 mg mL−1 insulin, 10% FBS, and 1% Pen/Strep. Cells were cultured in 75 cm2 tissue culture flask from TPP at 37 °C and 5% CO2 in a CO2 water jacketed incubator 3110 from Scientific Inc. Once weekly, the medium was changed and the trypsinization of confluent cells was performed with trypsin-ethylenediaminetet- raacetic acid solution from ATCC (#30-2101) for subculturing as recommended by the supplier. Cell counting was performed with a 2%-trypan-blue solution in PBS from VWR and dispos-able hemacytometers from Incyto C-chip.
Trypsin ethylenediaminetet raacetic acid solution
Manufactured by American Type Culture Collection
Sourced in United States
Trypsin-ethylenediaminetetraacetic acid (Trypsin-EDTA) solution is a cell culture reagent used to detach adherent cells from a culture vessel. It contains the proteolytic enzyme trypsin and the chelating agent EDTA, which together disrupt the cell-to-cell and cell-to-surface attachments, allowing the cells to be suspended for further passaging or analysis.
Automatically generated - may contain errors
Lab products found in correlation
Mcf 7, by American Type Culture Collection (2 mentions)
Axiovert 25, by Zeiss (2 mentions)
Alexa fluor 350, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Nile red, by Merck Group (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (1 mentions)
F 12k medium, by American Type Culture Collection (1 mentions)
L 15 medium, by American Type Culture Collection (1 mentions)
Trypan blue, by Merck Group (1 mentions)
Atp determination kit, by Thermo Fisher Scientific (1 mentions)
Cyquant nf cell proliferation assay kit, by Thermo Fisher Scientific (1 mentions)
Mitotracker deep red fm, by Thermo Fisher Scientific (1 mentions)
Rhodamine red x, by Thermo Fisher Scientific (1 mentions)
Fluorescein isothiocyanate (fitc), by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi, by Thermo Fisher Scientific (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by American Type Culture Collection (1 mentions)
Streptomycin, by American Type Culture Collection (1 mentions)
Scc15, by American Type Culture Collection (1 mentions)
4 protocols using trypsin ethylenediaminetet raacetic acid solution
1
Culturing HEK-293 and MCF7 Cell Lines
2
Culturing HEK-293 and MCF7 Cell Lines
3
Polymeric Nanoparticle Formulation Development
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PLGA
(Lactel 50:50, mol.
weight 30 000–60 000) was purchased from Durect
Corporation (Cupertino, CA). Chitosan (mol. weight 50 000–190 000)
was purchased from Sigma-Aldrich (St. Louis, MO). DOX and PTX were
ordered from LC Laboratories, Inc. (Woburn, MA, US). Coumarin-6,
fluorescein isothiocyanate, dichloromethane (DCM), polyvinyl alcohol
(PVA), and Nile red were purchased from Sigma-Aldrich, Inc. (Saint
Louis, MO). Ready-to-use dialysis tubes [molecular weight cut-off
(MWCO), 6000–8000] were purchased from Spectrum Laboratories,
Inc. (Rancho Dominguez, CA, US). 4′,6-Diamidino-2-phenylindole
(DAPI) and Alexa Fluor 350 were obtained from Life Technologies Corporation
(Grand Island, NY, USA). Cancer cell lines (MDA-MB-231 and A-549)
and related agents including trypsin/ethylenediamine tetraacetic acid
solution, F-12K medium, L-15 medium, and fetal bovine serum were purchased
from American Type Culture Collection (ATCC) (Manassas, VA, USA).
All of the other chemicals were of analytical grade.
(Lactel 50:50, mol.
weight 30 000–60 000) was purchased from Durect
Corporation (Cupertino, CA). Chitosan (mol. weight 50 000–190 000)
was purchased from Sigma-Aldrich (St. Louis, MO). DOX and PTX were
ordered from LC Laboratories, Inc. (Woburn, MA, US). Coumarin-6,
fluorescein isothiocyanate, dichloromethane (DCM), polyvinyl alcohol
(PVA), and Nile red were purchased from Sigma-Aldrich, Inc. (Saint
Louis, MO). Ready-to-use dialysis tubes [molecular weight cut-off
(MWCO), 6000–8000] were purchased from Spectrum Laboratories,
Inc. (Rancho Dominguez, CA, US). 4′,6-Diamidino-2-phenylindole
(DAPI) and Alexa Fluor 350 were obtained from Life Technologies Corporation
(Grand Island, NY, USA). Cancer cell lines (MDA-MB-231 and A-549)
and related agents including trypsin/ethylenediamine tetraacetic acid
solution, F-12K medium, L-15 medium, and fetal bovine serum were purchased
from American Type Culture Collection (ATCC) (Manassas, VA, USA).
All of the other chemicals were of analytical grade.
4
Cell Line Characterization and Culture Protocol
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BJ (ATCC-CRL 2522) and SCC-15 (ATCC-CRL-1623) cell lines and Dulbecco’s Modified Eagle’s Medium (DMEM-F12), Eagle’s minimum essential medium (EMEM), fetal bovine serum (FBS), penicillin, streptomycin, trypsin/ethylenediaminetetraacetic acid solution, and phosphate-buffered saline (PBS; with and without magnesium and calcium ions) were obtained from American Type Culture Collection (ATCC) (Manassas, VA, USA). Hydrocortisone, Trypan blue, and fluorescein isothiocyanate (FITC) were provided by Sigma-Aldrich Co. (St Louis, MO, USA). The fluorescent markers 4′,6-diamidino-2-phenylindole, dihydrochloride (DAPI), MitoTracker® Deep Red FM and Rhodamine Red™-X (succinimidyl ester, 5-isomer), ATP Determination Kit, and CyQUANT® NF Cell Proliferation Assay Kit were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Cell culture sterile plastics were purchased from Corning Incorporated (Corning, NY, USA), Sigma-Aldrich Co., and Brand GMBH + CO KG (Wertheim, Germany).
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