The ovarian cancer cell lines with a different clinical origin (32 (link),33 (link)): Caov-3 (ATCC® HTB-75), NIH:OVCAR-3 (ATCC® HTB-161, American Type Culture Collection) were cultured in the Dulbecco's Modified Eagle Medium (DMEM; Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and RPMI Medium-1640 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 20% FBS and 0.6 mg insulin (Insulin solution from bovine pancreas; Sigma-Aldrich; Merck KGaA)/500 ml medium, respectively. Cell line AAV-293 (Agilent Technologies) derived from human embryonic kidney cells was cultured in DMEM supplemented with 10% FBS. Human fibroblasts CCD-18Co (ATCC® CRL-1459) were cultured in Minimum Essential Media (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS. The media were also supplemented with 1% Antibiotic-Antimycotic (Gibco; Thermo Fisher Scientific, Inc.), and the cells were maintained at 37°C in a humidified 5% CO2 atmosphere. Mycoplasma test was performed in our laboratory, and its shown no contamination. The analyzed cells were verified in the International Cell Line Authentication Committee and ExPASy Cellosaurus databases (34 (link),35 (link)), in order to exclude their contamination with other cell lines or their incorrect identification.
Insulin solution from bovine pancreas
Manufactured by Merck Group
Sourced in United States
Insulin solution from bovine pancreas is a lab equipment product that provides a standardized source of insulin. It is extracted from the pancreas of cattle and supplied in a liquid form for use in various research and analytical applications. The product offers a consistent and reliable source of insulin for activities such as cell culture, enzymatic assays, and other experiments requiring insulin.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Hydrocortisone, by Merck Group (3 mentions)
Caov3, by American Type Culture Collection (1 mentions)
Nih ovcar3, by American Type Culture Collection (1 mentions)
Aav 293, by Agilent Technologies (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Heat inactivated fetal bovine serum, by Thermo Fisher Scientific (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Trypsin edta 0.25, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Caov3, by Thermo Fisher Scientific (1 mentions)
Skov3, by Thermo Fisher Scientific (1 mentions)
Collagenase type 1, by Merck Group (1 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (1 mentions)
Cls4520, by Corning (1 mentions)
Mcdb 105, by Merck Group (1 mentions)
Ovcar3, by Thermo Fisher Scientific (1 mentions)
11 protocols using insulin solution from bovine pancreas
1
Culturing Ovarian Cancer and Control Cell Lines
2
Culturing and Characterizing Human Breast Cancer BT474 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human breast cancer BT474 cell line (ATCC© HTB-20™) was obtained from American Type Culture Collection. The cells were maintained in DMEM/F-12 medium with GlutaMAX (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (Gibco), 5 μg/ml insulin solution from bovine pancreas (Sigma), 10 mM HEPES (Gibco), 100 U/ml penicillin, and 100 μg/ml streptomycin (Gibco). Cells were grown at 37°C under 5% CO2 in a humidified incubator. Prior to experimentation, cells (passages 8–18) were cultured in a 75 cm² flask to approximately 70 to 80% confluence. Cells were detached with 1X Trypsin-EDTA 0.25% (Gibco), and the expression of HER2 was monitored using an anti-CD340 (clone 24D2) and the corresponding isotype control (both from Miltenyi Biotec) using flow cytometry (data not shown).
3
Culturing Ovarian Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ovarian cancer cell lines were purchased from ATCC® (LGC Ltd). SKOV3 (CVCL_0532) and OVCAR-3 (CVCL_0465) were maintained in RPMI 1640 media (Gibco™, 11875093) supplemented with 20% foetal calf serum (FBS; Gibco™, 10270106) and 10 μg/ml of insulin solution from bovine pancreas (Sigma, I0516). UWB1.289 (BRCA1mut; CVCL_B079) and CAOV3 (CVCL_0201) were maintained in DMEM/F-12 + GlutaMAX™ (Thermo Scientific, 31331093) (10% FBS). None of the cell lines we have used are frequently misidentified. Patient derived primary ovarian cancer cells were isolated from ovarian biopsies using a protocol adapted from Shepherd et al. [64 (link)]; the main difference is the use of Collagenase type I (2 mg/mL; Sigma, 17100–017) instead of Dispase II. Primary cells were maintained in 50% MCDB 105 (Sigma, M6395) + 50% M199 (Gibco, 31150–022) (20% FBS). All media was supplemented with penicillin [100 U/ml] and streptomycin [100 μg/ml] (P/S; Gibco, 15140–122), and cells were maintained at 37 °C in a humidified 5% CO2 incubator. 3D spheroids were grown in 96-well ultra-low attachment (ULA) plates (Corning, CLS4520). The human biological samples were sourced ethically, and their research use was in accord with the terms of the informed consents under an IRB/EC approved protocol. Mycoplasma contamination was routinely tested with the MycoAlert Mycoplasma Detection kit (Lonza, LZLT07218).
4
MCF7 Breast Cancer Cell Culturing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MCF7 cells, a breast cancer cell line of the luminal type (Figure S1 ), were derived from a single progenitor cell and were grown in DMEM (Dulbecco's Modified Eagle Medium) with 4.5 g/L glucose and L-Glutamine, without sodium pyruvate (DMEM, Corning), supplemented with 10% fetal bovine serum (FBS, Atlanta Biologicals), penicillin (100 U/mL, Gibco) and streptomycin (100 mg/mL, Gibco), and insulin at 10 μg/ml (Insulin solution from bovine pancreas, Sigma). The cells were grown to 80% confluency. They were passaged approximately every 3-4 days: washed with 1XPBS (PBS, pH 7.4, Gibco), trypsinized (Trypsin-EDTA, Gibco) and split at about 1:4 ratio).
5
Melanoma Spheroid Formation Assay
6
Glucose and Insulin Response in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were fasted for 12 hours or 2 hours and then injected subcutaneously with glucose (2 g/kg body weight) or insulin (0.5 U/kg body weight; insulin solution from bovine pancreas, #I0516; Sigma‐Aldrich, St. Louis, Missouri), respectively, with a rest period of 5 days between tests. Blood samples were taken every 20 minutes (0‐120 minutes) from the tail vein, and blood glucose levels were determined with a portable glucometer (Accu‐Chek Aviva 05911982002; Roche Diagnostics, Basel, Switzerland). The area under the curve was calculated as the sum of trapezoids in both tests.
7
Chondrogenic Differentiation of ATDC5 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ATDC5 cells were kindly provided by Professor Chisa Shukunami (Hiroshima University, Hiroshima, Japan). The cells were maintained in Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (DMEM/F-12) (Gibco, New York, NY) containing 5% fetal bovine serum (Sigma, St. Louis, MO) (13 (link), 14 (link)). The cells were plated at a density of 5.0 × 104 cells/cm2 in a 6-well plate for RNA-Seq and quantitative real-time reverse transcription polymerase chain reaction (RT-PCR), a 10-cm dish for western blotting, and a 24-well glass bottom plate (Iwaki, Tokyo, Japan) for imaging analysis. For induction of chondrogenesis, the cells were cultured in a differentiation medium which was composed of DMEM/F-12 containing 5% fetal bovine serum, 10 μg/mL insulin solution from bovine pancreas (Sigma-Aldrich, St. Louis, MO), 10 μg/mL human apo-transferrin (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan), and 3 × 10–8 M sodium selenite (FUJIFILM Wako Pure Chemical Corporation) from the cell seeding.
8
3D Mammary Epithelial Acini Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HMT3522 T4-2 (V. Weaver, UCSF) cells were cultured in precoated collagen plates using DMEM/Ham's F12 (1:1) medium supplemented with 2 mM glutamine (Life Technologies), 250 ng/mL insulin solution from bovine pancreas (Sigma-Aldrich), 10 µg/mL transferrin (Sigma-Aldrich), 2.6 ng/mL sodium selenite (Sigma-Aldrich), 10−10 M 17 β-estradiol (Sigma-Aldrich), 1.4 × 10−6 M hydrocortisone (Sigma-Aldrich), and 10 ng/mL human prolactin (Miltenyi Biotec).
3D acini were grown as follows: single-cell suspensions (1.5 × 104 cells per mL) were plated in the appropriate medium supplemented with 2% Growth Factor Reduced Matrigel (GFRM; BD Biosciences). 100 μL of this mix were added per well in a 96-well ImageLock plate (Essen Biosciences) precoated with 10 μL of pure GFRM for 15 min at 37°C. Cells were incubated at 37°C for 5 days, changing the media every two days, before IF.
For inhibitor studies, cells were treated from the time of plating with Tyrphostin-AG1478 (80 nM in ethanol, Sigma-Aldrich), erlotinib (100 nM in DMSO), and ethanol or DMSO as appropriate controls, respectively.
HEK293-FT (Thermo Fisher Scientific) were cultured in DMEM supplemented with 10% FBS, 6 mM L-glutamine, and 0.1 mM non-essential amino acids (NEAA) (all reagents from Life Technologies/Thermo Fisher).
3D acini were grown as follows: single-cell suspensions (1.5 × 104 cells per mL) were plated in the appropriate medium supplemented with 2% Growth Factor Reduced Matrigel (GFRM; BD Biosciences). 100 μL of this mix were added per well in a 96-well ImageLock plate (Essen Biosciences) precoated with 10 μL of pure GFRM for 15 min at 37°C. Cells were incubated at 37°C for 5 days, changing the media every two days, before IF.
For inhibitor studies, cells were treated from the time of plating with Tyrphostin-AG1478 (80 nM in ethanol, Sigma-Aldrich), erlotinib (100 nM in DMSO), and ethanol or DMSO as appropriate controls, respectively.
HEK293-FT (Thermo Fisher Scientific) were cultured in DMEM supplemented with 10% FBS, 6 mM L-glutamine, and 0.1 mM non-essential amino acids (NEAA) (all reagents from Life Technologies/Thermo Fisher).
9
SV40-Immortalized Bovine Mammary Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BIE cells [17 (link)], which were immortalized by the
simian virus 40 large T antigen (SV40 T) gene, were kindly provided by Dr. Aso (Tohoku
University, Sendai, Japan) and maintained in DMEM (044-29765, Wako) containing 4,500
mg/l glucose, 100 U/ml penicillin, 100
µg/ml streptomycin (15140-122, Gibco, Grand Island,
NY, USA), and 10% FBS. SV40 T-immortalized MAC-T cells [9 (link)] were kindly provided by Dr. Roh (Tohoku University) and maintained in DMEM
(041-29775, Wako) containing 1,000 mg/l glucose, 100
U/ml penicillin, 100 µg/mlstreptomycin, 10% FBS, 10 mg/ml insulin solution from bovine pancreas
(10516, Sigma, St. Louis, MO, USA), and 10 mg/ml hydrocortisone (H0888,
Sigma). MAC-T cells are known to secret α- and β-casein [9 (link)], indicating that MAC-T cells are derived from the mammary glandular
epithelium.
simian virus 40 large T antigen (SV40 T) gene, were kindly provided by Dr. Aso (Tohoku
University, Sendai, Japan) and maintained in DMEM (044-29765, Wako) containing 4,500
mg/l glucose, 100 U/ml penicillin, 100
µg/ml streptomycin (15140-122, Gibco, Grand Island,
NY, USA), and 10% FBS. SV40 T-immortalized MAC-T cells [9 (link)] were kindly provided by Dr. Roh (Tohoku University) and maintained in DMEM
(041-29775, Wako) containing 1,000 mg/l glucose, 100
U/ml penicillin, 100 µg/mlstreptomycin, 10% FBS, 10 mg/ml insulin solution from bovine pancreas
(10516, Sigma, St. Louis, MO, USA), and 10 mg/ml hydrocortisone (H0888,
Sigma). MAC-T cells are known to secret α- and β-casein [9 (link)], indicating that MAC-T cells are derived from the mammary glandular
epithelium.
10
Cell culture media and cytotoxicity assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Minimum essential medium Eagle alpha modification (α-MEM), Dulbecco’s modified Eagle’s medium (DMEM), Ham’s F-12 nutrient mixture medium (F12), penicillin–streptomycin solution, insulin solution from bovine pancreas, hydrocortisone, cholera toxin, human epidermal growth factor, gelatin, fibronectin and Dox were obtained from Sigma-Aldrich (St. Louis, MO, USA). Horse and fetal bovine sera (FBS) and trypsin/ethylenediamine tetraacetic acid (EDTA) solution were obtained from Gibco® (Thermo Fisher Scientific, Waltham, MA, USA). 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethonyphenol)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) was purchased from Promega (Madison, WI, USA). An 800 µM stock solution of doxorubicin hydrochloride (Dox) was prepared in H2O MilliQ, aliquoted and stored at −20 °C until further use.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!