Rat HSC-T6 and human LX-2 cell lines were obtained from Dr. Yun-Lian Lin at The National Research Institute of Chinese Medicine, Taiwan. HSC-T6 cells, immortalized early active stage rat hepatic stellate cells (HSCs), were maintained in Waymouth MB 752/1 medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal calf serum and antibiotics (100 IU/mL each of penicillin and streptomycin) in a humidified atmosphere containing 5% CO2 and 95% air at 37 °C. LX-2, immortalized early active stage human HSCs were maintained in Dulbecco’s Modified Eagle medium (DMEM) supplemented with 2% fetal calf serum and antibiotics (100 IU/mL each of penicillin and streptomycin) in a humidified atmosphere containing 5% CO2 and 95% air at 37 °C. The cultures were passaged by trypsinization every 3–4 days.
Waymouth mb 752 1 medium
Manufactured by Merck Group
Sourced in United States
Waymouth MB 752/1 medium is a cell culture medium formulated for the growth and maintenance of a variety of cell lines. It is a complete, defined, and serum-free medium that provides the necessary nutrients and growth factors to support cell proliferation and viability.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Heat inactivated fetal bovine serum, by Merck Group (2 mentions)
Dmem medium, by Corning (2 mentions)
Mycoplasma detection kit, by InvivoGen (2 mentions)
Erk1 2, by Cell Signaling Technology (1 mentions)
Sapk jnk, by Cell Signaling Technology (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Dulbecco s phosphate buffered saline, by Thermo Fisher Scientific (1 mentions)
Tween 20, by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Isopropyl alcohol, by Merck Group (1 mentions)
Gentamicin sulfate, by Thermo Fisher Scientific (1 mentions)
Sodium chloride (nacl), by Honeywell (1 mentions)
Paclitaxel, by Merck Group (1 mentions)
11 protocols using waymouth mb 752 1 medium
1
Immortalized Hepatic Stellate Cell Lines
2
Apoptosis Induction in Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cordycepin, paclitaxel, cisplatin, Waymouth MB 752/1 medium, propidium iodide, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), RNase A, bovine serum albumin, and dimethyl sulfoxide (DMSO) were purchased from Sigma Chemicals (St Louis, MO, USA). Fetal bovine serum, lyophilized trypsin-ethylenediaminetetraacetic acid (EDTA), Dulbecco’s phosphate-buffered saline (PBS), and gentamicin sulfate were purchased from Gibco (Grand Island, NY, USA). Sodium hydroxide, hydrochloric acid, sodium dodecyl sulfate, EDTA, isopropyl alcohol, chloroform, and Tween-20 were purchased from Merck (Darmstadt, Germany). Acrylamide was purchased from JT Baker (Phillipsburg, NJ, USA). HEPES was purchased from Mallinckrodt Baker Inc (Paris, KT, USA). Sodium bicarbonate, sodium carbonate, and sodium chloride were purchased from Riedel-de Haën (Seelze, Germany). Antibodies against cleaved caspase-3, caspase-8, and caspase-9, phospho-ERK1/2, phosphor-SAPK/JNK, phosphor-p38, ERK1/2, SAPK/JNK, and p38 were purchased from Cell Signaling (Beverly, MA, USA). Tris base, Z-LEHD-FMK (caspase-9 inhibitor), Z-IETD-FMK (caspase-8 inhibitor), Z-DEVD-FMK (caspase-3 inhibitor), and β-actin were purchased from Calbiochem (San Diego, CA, USA). Anti-PARP antibody was purchased from Oncogene (San Diego, CA, USA).
3
MLTC-1 Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MLTC-1 cells purchased from ATCC (Manassas, USA) were cultured in a T-75 flask in Waymouth MB 752/1 medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% heat-inactivated fetal bovine serum (Sigma-Aldrich) at pH 7.3 at 37°C in 5% CO2 in a humidified incubator. All experiments for this study were conducted on MLTC-1 cells between passage numbers 10 and 30. The number of cells to be used in experiments was standardized and was seeded at a density of 2 × 105 cells/ml/well for estimating steroid and VEGF production.
4
MLTC-1 Cells for Steroid Production
5
Maintenance of MA-10, TM4, and NT2/D1 Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MA-10 cells were a gift from Dr. Mario Ascoli (University of Iowa, Iowa City, IA)48 (link). Cells were maintained in Waymouth MB 752/1 medium (Sigma-Aldrich, St. Louis, MO) containing 10% fetal bovine serum (Invitrogen, Grand Island, NY) and incubated in a humidified atmosphere containing 95% air and 5% CO2 at 37 °C. The murine TM4 Sertoli-like cell line and NT2/D1 cells were obtained from the American Type Culture Collection (Manassas, VA)49 (link). TM4 Sertoli cells were maintained in Dulbecco’s modified eagle medium, nutrient mixture F-12 (DMEM/F-12; Invitrogen, Grand Island, NY) containing 10% fetal bovine serum and incubated in a humidified atmosphere containing 95% air and 5% CO2 at 37 °C. NT2/D1 cells were maintained in Dulbecco’s modified eagle medium with high glucose (DMEM/HIGH GLUCOSE; Invitrogen, Grand Island, NY) containing 10% fetal bovine serum and incubated in a humidified atmosphere containing 95% air and 5% CO2 at 37 °C.
6
Culturing Immortalized Rat Hepatic Stellate Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HSC-T6, a generous gift from Prof. S.L. Friedman, is an immortalized cell line of rat HSCs [4] (link). HSC-T6 cells were incubated in Waymouth MB 752/1 Medium (Sigma-Aldrich, St. Louis, MO, USA) containing 10% fetal bovine serum (FBS, pH 7.0; Gibco BRL, Gaithersburg, MD, USA) at 37°C in 5% CO2.
7
Characterization of 1000W Rat Tracheal Cell Line
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 1000W cell line was derived as an outgrowth from a heterotopic tracheal transplant in a F344 inbred rat, following exposure to 7,12‐dimethylbenz(a)anthracene (Marchok, Rhoton, & Nettesheim, 1978 ). The epithelial cells were cultured in Waymouth MB752/1 medium (Sigma–Aldrich, St. Louis, MO) supplemented with 10% fetal bovine serum (Hyclone, UT or Atlanta Biologicals, GA), as previously described (Amarachintha et al., 2015 ; Heckman, Plummer, & Runyeon, 1996 ). The line was tested periodically and found to be free of contamination with Mycoplasma. The 1000‐W cells were tested periodically for tumorigenicity in immune‐suppressed, syngeneic animals. After 65 weeks in culture, they were positive for tumorigenicity, suggesting that long‐term culture in vitro has an effect similar to tumor promotion. Cells used in these experiments were at 35–45 weeks of in vitro culture. For experiments, glass coverslips were coated with germanium for use as substrates (see Preparation of substrates). Cells were subcultured at a density of 250,000 per 35‐mm dish and incubated overnight to allow them to adhere to the substrates. Then, they were treated with reagents as described below (see Treatment with BPs, small inhibitory RNAs (siRNAs), and ODNs).
8
Establishment and Maintenance of KPC and KPYC Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
KPC/vector and KPYC/Flag-Yap lines were established previously and cultured on collagen-coated plates in Waymouth MB 752/1 Medium (Sigma-Aldrich, St. Louis, MO) containing 10% FBS (Zhang et al 2014 (link)). KPC cells established from a KPC PDAC tumor were stably transduced with pTRIPZ or pTRIPZ-Yapsh and maintained in DMEM medium (Corning, Corning, NY) containing 10% fetal bovine serum (FBS). Yapsh expression was induced by treating the cells with Doxcycline (4 µg/mL) for 3 days. Regularly mycoplasma testing was performed in house using Mycoplasma Detection Kit (InvivoGen, San Diego, CA).
9
Isolation and Culture of Murine Pancreatic Cells
10
Establishment and Maintenance of KPC and KPYC Cell Lines
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!