MCF7 cells were sourced from the Michigan Cancer Foundation. Derivatives of MCF7 cells that are resistant to tamoxifen (TamR) or long-term estrogen deprived (LTED) were obtained from Myles Brown (Dana-Farber Cancer Institute) (Bailey et al. 2015) (link). T47D cells were obtained from the American Type Culture Collection. All cell lines have been verified through short tandem repeat profiling, and mycoplasma contamination was excluded. MCF7, TamR and T47D cell lines were cultured in RPMI 1640 media (Thermo Fisher) supplemented with 10% fetal bovine serum (GE Healthcare), 20 mM HEPES (Thermo Fisher) and 0.28 IU/mL insulin (Novo Nordisk). Base media for TamR cells was also supplemented with 5 µM 4-hydroxytamoxifen (Sigma). TamR cells used in experiments were grown in base media without tamoxifen. The LTED cell line was maintained in phenolred free RPMI 1640 media (Thermo Fisher) supplemented with 10% steroid-depleted FBS (Thermo Fisher), 20 mM HEPES (Thermo Fisher) and 0.28 IU/mL insulin (Novo Nordisk). All cells were cultured in a humidified incubator at 37°C with 5% CO 2 and were used at <10 passages post revival. The AR antagonist enzalutamide (Selleck) was re-suspended as a 40 mM stock solution in DMSO and DHT (Sigma) was re-suspended as a 10 µM stock solution in ethanol.
Insulin
Manufactured by Novo Nordisk
Sourced in Denmark, United States, China, Germany, United Kingdom, Switzerland, France, Canada
Insulin is a laboratory product used for research and scientific experimentation. It is a hormone produced by the pancreas that regulates blood sugar levels. Insulin plays a crucial role in the metabolism of carbohydrates, fats, and proteins in the body.
Automatically generated - may contain errors
Lab products found in correlation
Dexamethasone (dex), by Merck Group (13 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (11 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Dmem f12, by Thermo Fisher Scientific (6 mentions)
Hydrocortisone, by Merck Group (5 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Epidermal growth factor (egf), by Merck Group (4 mentions)
Mcf 7, by American Type Culture Collection (4 mentions)
D glucose, by Novo Nordisk (3 mentions)
Glucometer, by Roche (3 mentions)
Accu chek, by Roche (3 mentions)
One touch ultra 1, by LifeScan (3 mentions)
Free style, by Nipro (3 mentions)
Pyruvate, by Merck Group (3 mentions)
3 isobutyl 1 methylxanthine, by Merck Group (3 mentions)
Mda mb 231, by Thermo Fisher Scientific (3 mentions)
Gentamicin, by Thermo Fisher Scientific (3 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (3 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (3 mentions)
160 protocols using insulin
1
Establishing Tamoxifen-Resistant and Estrogen-Deprived Breast Cancer Cell Lines
2
Differentiation and Treatment of Murine Adipocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The freshly collected BAT between the scapulae region of the mice was washed with pre-chilled high-sugar DMEM (Gibco, USA). Tissues were cut, digested, then filtered, centrifuged, and suspended with DMEM containing 20% fetal bovine serum (FBS, Gibco, USA) and 1% penicillin/streptomycin (P/S, sigma, USA). Cells were plated, cultured, and induced to differentiate according to our previously reported procedures (17 (link)). The initial differentiation medium was supplemented with 0.5 mM isobutylmethylxanthine, 0.5 μM dexamethasone, 0.125 mM indomethacin, 1 nM T3 (Sigma, USA), and 20 nM insulin (Novo Nordisk, China) for 48 hours and then replaced by differentiation medium containing the same concentration of T3 and insulin for further culture until 5-6 days. For treatment, Forskolin (HY-15371, MedChemExpress Inc., China) was added to preadipocytes and differentiated adipocytes were cocultured with M2 macrophages.
3
Comparative Insulin Regimen Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Using an independently-generated randomization list, patients were consecutively assigned to one of two insulin regimens after enrollment: AI (insulin aspart and insulin detemir; insulin by Novo Nordisk, Bagsvaerd, Denmark) or HI (regular insulin and NPH; insulin by Novo Nordisk, Bagsvaerd, Denmark). An open label design was used as the administration time of the meal-related insulin differed between the two insulin regimens. However, any staff involved in ultrasound investigations, blood tests or the definition of data sets for statistical analysis remained blinded until the database was locked.
4
Hyperinsulinemic-Euglycemic Clamp Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
VATP was assessed during hyperinsulinemic-euglycemic conditions as shown in Fig. 3B . At the beginning of this study, we infused a bolus of 5 mU/kg/min of insulin, followed by 3 mU/kg/min of insulin (Novo Nordisk, Bagsværd, Denmark) at a rate of 2.1 μl/min and 12.5 mg/kg/min of 20% dextrose to maintain euglycemia during hyperinsulinemia. Blood samples were taken retro-orbitally after the clamp. At study completion, mice were anesthetized and tissues were harvested within 3 min with liquid N2–cooled aluminum tongs and stored at –80°C for subsequent analysis.
5
Metabolic Phenotyping in Murine Models
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Oral glucose tolerance test (OGTT) The mice were fasted for 14 h and then gavaged glucose (2 g/kg). At 0, 15, 30, 60, 90, and 120 min after gavage the tail vein blood glucose concentration was measured.
Insulin tolerance test (ITT) The mice were fasted for 4 h and then intraperitoneally injected with Insulin (0.75 IU/kg; Novo Nordisk, Beijing, China). At 0, 15, 30, 45, 60, and 90 min after intraperitoneal injection the tail vein blood glucose concentration was measured.
Determination of serum Insulin content The serum Insulin level was calculated according to the instructions of the ELISA kit (H203‐1‐2; Njjcbio, Nanjing, China).
Homeostasis model evaluation method‐Insulin resistance index (HOMR‐IR) This was calculated as follows [HOMR‐IR = fasting serum Insulin (mIU/L)*fasting blood glucose (mmol/L)/22.5].
Insulin tolerance test (ITT) The mice were fasted for 4 h and then intraperitoneally injected with Insulin (0.75 IU/kg; Novo Nordisk, Beijing, China). At 0, 15, 30, 45, 60, and 90 min after intraperitoneal injection the tail vein blood glucose concentration was measured.
Determination of serum Insulin content The serum Insulin level was calculated according to the instructions of the ELISA kit (H203‐1‐2; Njjcbio, Nanjing, China).
Homeostasis model evaluation method‐Insulin resistance index (HOMR‐IR) This was calculated as follows [HOMR‐IR = fasting serum Insulin (mIU/L)*fasting blood glucose (mmol/L)/22.5].
6
Glucose and Insulin Tolerance Assays in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For OGTT, mice were fasted overnight while having access to drinking water and administered with glucose solution (Sigma-Aldrich) via oral gavage (2 g glucose/kg body weight). Blood samples were collected (∼30 μL) at the indicated time points of glucose administration and used for measuring glucose levels with a glucometer and Insulin levels by Ultrasensitive Mouse Insulin ELISA kit (Crystam Chem). For ITT, mice were fasted for 2 h before Insulin injection while ensuring that the mice had access to drinking water. Insulin (Novo Nordisk) was injected intraperitoneally (0.35 to 0.75 U Insulin/kg body weight). Blood glucose levels were measured before and after the indicated time points of Insulin injection.
7
Glucose and Insulin Tolerance Tests
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Intraperitoneal glucose tolerance test was conducted at the end of the dietary intervention period by intraperitoneal administration of 2.5 g glucose per kg body weight.[41 ] The animals were fasted for 6 h prior to the test. For intraperitoneal insulin tolerance tests, animals were fasted for 4 h prior to the intraperitoneal injection of insulin (Novo Nordisk, Denmark) at 0.75 unit kg−1 body weight.
8
Glucose and Insulin Tolerance Tests
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were fasted for 6 h prior to glucose tolerance test and 4 h prior to Insulin tolerance test, starting at 6:00 a.m. Mice were single-housed during the fasting period and the duration of the test. Water was provided ad libitum. Basal glucose levels were measured before intraperitoneal injection of the glucose or Insulin solution (fixed dose of 42 mg glucose (Sigma-Aldrich, St. Louis, MI, United States), and 0.02 IU Insulin (Novo Nordisk, Bagsvaerd, Denmark)), respectively. Blood glucose levels (mg/dl, measured with Accucheck guide glucometer (Roche, Basel, Switzerland)) were measured after 15, 30, 60, and 120 min of glucose injection. Blood samples were taken from the tail vein.
9
Intraperitoneal Glucose and Insulin Tolerance Tests
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the intraperitoneal glucose tolerance test (IPGTT) and insulin tolerance test (ITT), mice were fasted overnight and then given an intraperitoneal injection of D-glucose (1.5 g/kg) or insulin (0.75 UI/kg; Novo Nordisk). Blood samples were harvested from the tail vein at different time points, and the blood glucose was measured using a glucometer (Johnson & Johnson, Shanghai, China). After testing serum fasting blood glucose and insulin concentrations, we calculated the homeostasis model index of insulin resistance (HOMA-IR: insulin × glucose/22.5) and the quantitative insulin sensitivity check index (QUICKI: 1/[log (insulin) + log (glucose)]), as previously described [49 (link)].
10
Isolation and Culture of Gastric Cancer Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human gastric cancer cell lines (SGC7901 and MKN45) were purchased from the cell bank of the Chinese Academy of Sciences (Shanghai, China). Cells were frozen at low passage and used within 2–3 months after thawing. SGC7901 cells and MKN45 cells were cultured in DMEM (Gibco, USA) and RPMI-1640 (Gibco, USA), supplemented with 10% FBS, respectively. Human gastric cancer stem cell lines (SGC-CSC and MKN-CSC) were isolated from SGC7901 and MKN45 cells by performing a sphere-forming culture. In short, the GC cells were plated at a density of 104 cells/well in ultra-low-attachment 6-well plates (Corning) with stem cell medium, comprised GlutaMAX-DMEM/F12 (Gibco, USA), 2% B27 (Invitrogen), epidermal growth factor (20 ng/mL, Peprotech), basic fibroblast growth factor (10 ng/mL, Peprotech), 0.4% bovine serum albumin (Solarbio), and 5 μg/mL insulin (Novo Nordisk). Culture medium was supplemented with additional growth factors every 3 days, and the cells were maintained in stem cell medium for 7 days. Tumorspheres were collected, dissociated into single-cell suspensions, and cultured to acquire the regeneration of tumorspheres. Second-passage tumorspheres were used for all relevant experiments. All cells were maintained at 37 °C in a 5% CO2 humidified atmosphere and tested for mycoplasma contamination before 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!