The plant material (leaves, branches, and fruits) from Inga species (Inga laurina, Inga marginata, and Inga edulis) were dried at room temperature, ground, and extracted with ethanol for 20 min, using an ultrasonic bath. After evaporation under reduced pressure using a rotary evaporator (54-Rotavapor R-220, brand: Büchi, with vacuum pump Vacuum Controller V-805, brand: Büchi and water circulator), the EtOH extracts were submitted to cytotoxicity evaluation against HT-29 human colon cancer cells (ATCC® HB-8248™, Manassas, VA, USA) and T98G human glioblastoma cancer cell lines (ATCC® CRL-1690™), using doxorubicin as a positive control.
Crl 1690
Manufactured by American Type Culture Collection
Sourced in United States
CRL-1690 is a cell line derived from human embryonic kidney cells. It is a commonly used cell line for various research purposes, including the study of gene expression, protein production, and cell signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
54 rotavapor r 220, by Büchi (1 mentions)
Htb 14, by American Type Culture Collection (1 mentions)
Gbm cell lines, by American Type Culture Collection (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Avantor (1 mentions)
Recombinant human egf, by Thermo Fisher Scientific (1 mentions)
Htb 15, by American Type Culture Collection (1 mentions)
Mycoalert system, by Cambrex (1 mentions)
Pta 5077, by American Type Culture Collection (1 mentions)
Ccl 107, by American Type Culture Collection (1 mentions)
Tc plate 24 well, by Greiner (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
U87mg, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomicin 100x, by Euroclone (1 mentions)
Glucose assay kit, by Abcam (1 mentions)
Htb 96, by Merck Group (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Ln229, by Thermo Fisher Scientific (1 mentions)
Exosome depleted fbs, by Thermo Fisher Scientific (1 mentions)
10 protocols using crl 1690
1
Cytotoxicity Evaluation of Inga Species
2
Cultivation of Human Glioblastoma Cell Lines
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All cells used in the present study were human glioblastoma multiforme (GBM) cell lines, purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). These cell lines were Ln18 (ATCC®, CRL-2610™), U87 (ATCC®, HTB-14™), T98G (ATCC®, CRL-1690™), and M059K (ATCC®, CRL-2365™). All cells were grown in RPMI 1640 without phenol red, supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, and penicillin (100 IU mL−1) /streptomycin (100 μg mL−1) at 37 °C in a 5% CO2 and 95% humidified atmosphere.
3
Synchronized Cell Cycle Phases in T98G Glioblastoma Cells
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T98G cells (origin human glioblastoma multiforme, ATCC® CRL-1690™) were cultured at 37 °C and 7% CO2 in DMEM supplemented with 10% serum and 2 mM l -glutamine. Cells were synchronized in G1 using serum starvation for 72 h and S-phase cells were acquired by serum starving cells for 2 days, followed by a 22 h release in 10% serum, as described previously72 (link). Mitotic cells were synchronized as previously described73 (link) with slight modifications. Cells were first starved for 48 h, and then released into 20% serum containing media supplemented with 0.2 μg/mL nocodazole for 36 h. Mitotic cells were subsequently collected by shake-off.
4
Differentiated GBM Cell Lines and Glioma Stem Cells
5
Evaluation of Nanoparticle Cytotoxicity
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The influence of the Au-CuO and CuO-ZnO nanoparticles compounds on the viability of cells in vitro was evaluated with use two established cell lines received from American Type Culture Collection (Manassas, VA, USA). The rat brain glioma cell line C6 (ATCC® CCL-107™) was cultured in DMEM medium containing 10% fetal bovine serum, 50 U-mL penicillin, 100 μg-mL streptomycin, and 25 μg-mL amphotericin B. The human glioblastoma cell line T98G (ATCC® CRL-1690™) was grown in EMEM medium supplemented as mentioned above. Cells were kept at 37 °C in a 5% CO2 atmosphere in the incubator with increased humidity up to 85% confluency. Cells were plated in quantity 5 × 105 cells per well and cultured in the plastic 24-Well flat-bottom plates (TC-PLATE 24 well, Greiner).
6
Cultivating Human GBM Cell Lines
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Three human GBM cell lines, A172 (CRL-1620™; ATTC, Manassas, VA, USA), U87MG (HTB-14™; ATTC) and resistant-T98G (CRL-1690™; ATCC) were obtained. U87MG and resistant-T98G were cultured in Eagle's minimum essential medium (Thermo Fisher Scientific, Inc., Waltham, MA, USA), while A172 cells were grown in Dulbecco's modified Eagle's medium (Thermo Fisher Scientific, Inc.). Each medium was supplemented with 10% fetal bovine serum (cat. no. ECS0180L; EuroClone SpA, Via Figino, Milan, Italy) and 1% penicillin-streptomycin (Penicillin/Streptomicin 100X; cat. no. ECB3001D; EuroClone SpA). All cell lines were maintained in a humidified atmosphere of 5% CO2/95% air at 37°C. Cells from exponentially growing cultures were used for all experiments.
7
Simvastatin Effect on Glucose Metabolism in T98G Cells
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T98G cells (ATCC® CRL1690™) were grown in DMEM high glucose, supplemented with 10% FCS, sodium pyruvate (1 × 10−3m ), penicillin (100 U mL−1), and streptomycin (100 µg mL−1) at 37 °C and 5% CO2 and seeded in 12‐well plates (6 × 104 cells per well). After 24 h, medium was changed to phosphate buffered saline (PBS) containing 0.1% DMSO and simvastatin (1 × 10−6, 6 × 10−6, 10 × 10−6m ) or 0.1% DMSO alone and cells were incubated at 37 °C and 5% CO2 for 2 h. Medium was then changed to 850 µL of a 4:1 mixture of PBS and DMEM low glucose medium supplemented with 10% FCS, sodium pyruvate (1 × 10−3m ), penicillin (100 U mL−1), and streptomycin (100 µg mL−1) additionally containing 0.1% DMSO and simvastatin (1 × 10−6, 6 × 10−6, 10 × 10−6m ) or 0.1% DMSO alone. 50 µL of supernatant was then sampled at time points 0, 3, 18, and 24 h, centrifuged at 14 000 × g for 5 min, snap‐frozen in liquid nitrogen and stored at −80 °C until further use. Glucose content in the sampled supernatants was quantified using a fluorometric assay (Abcam glucose assay kit #ab65333, Abcam, Cambridge, United Kingdom), following the manufacturer's instructions.
8
Culturing Glioblastoma, Astrocyte, and Osteosarcoma Cells
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The T98G glioblastoma multiforme cell line (ATCC CRL-1690) was obtained from ATCC and cultured based on the specifications provided by ATCC. The human astrocyte cell line (ScienCell 1800) was purchased from ScienCell and was similarly cultured based upon the directions provided by ScienCell. GM00637 normal human fibroblasts, SV-40 transformed (Coriell) and U2OS human bone osteosarcoma epithelial cells (ATCC HTB-96) were cultured in DMEM (Sigma D5796) containing 10% FBS (Gemini 100-106) and 1% penicillin/streptomycin (Gibco 15070063).
9
Comparative Analysis of Glioma Cell Lines
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The SB28 and GL261 cell lines were purchased from the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ) with catalogue numbers ACC 880 and ACC 802. The HEK293T, MEF, U138, T98G, and LN-229 cell lines were purchased from the American Type Culture Collection (ATCC, CRL-3216, SCRC-1040, HTB-16, CRL-1690, and CRL-2611). The CT-2A, U87, and U251 cell lines were purchased from Millipore Sigma Aldrich with catalogue numbers SCC194, 89081402-1VL, and 09063001-1VL. HEK293T, MEF, GL261, LN229, and CT-2A cells were cultured in Dulbecco’s modified essential medium (DMEM) containing 10% heat-inactivated foetal bovine serum (FBS, Thermo Fisher Scientific, 26140095; Exosome-depleted FBS, Thermo Fisher Scientific, A2720801) supplemented with 1% penicillin/streptomycin at 37 °C in a humidified condition equilibrated with 5% CO2. SB28 cells were cultured in DMEM with 10% FBS supplemented with 1% penicillin–streptomycin at 37 °C and 10% CO2. U87, U251, U138, and T98G cells were cultured in Eagle’s minimum essential medium (EMEM) with 10% FBS supplemented with 1% penicillin–streptomycin at 37 °C and 5% CO2.
10
Glioma Cell Lines and Normal Controls
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The glioma tumour cell panel (T98G, A172 and SW1088) was purchased from American Type Culture Collection (ATCC) under catalogue numbers: CRL-1690™, CRL-1620™, HTB-12™; respectively). Cell cultures were maintained as monolayer in Dulbecco’s modified Eagle’s media with 25 mM glucose, and supplemented with foetal bovine serum (10%, v/v), and penicillin/streptomycin (1×; PAA). Normal Human Astrocytes (NHA) were provided from ATCC and cultured in Dulbecco’s modified Eagle’s media high glucose/F12 (1:1; Merck KGaA, Darmstadt, Germany) supplemented with foetal bovine serum (10%, v/v), and penicillin/streptomycin (1×; PAA). The Human Dermal Fibroblasts (HDFa; Gibco—Thermo Fisher Scientific, Waltham, MA, USA) was used as non-specific tissue control and cultured in HAM´s Nutrient Mixture F12 (Merck KGaA, Darmstadt, Germany) supplemented with foetal bovine serum (10% v/v; Gibco—Thermo Fisher Scientific, Waltham, MA, USA) and penicillin/streptomycin (1×; PAA Laboratories GmbH, Austria). Cells were cultured at 37 °C in an atmosphere of 5% CO2. Before each experiment, single-cell suspension was prepared using 0.05% trypsin/EDTA solution, and cells were counted using CountessTM automated cell counter (Thermo Fisher Scientific, Waltham, MA, USA).
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