The largest database of trusted experimental protocols

Mouse egf

Manufactured by Merck Group
Sourced in Germany, United States

Mouse EGF is a laboratory reagent that contains the epidermal growth factor (EGF) protein derived from mouse cells. EGF is a signaling molecule that plays a role in cell growth and proliferation. This product can be used in various cell culture and research applications.

Automatically generated - may contain errors

13 protocols using mouse egf

1

Isolation and Culture of Neural Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
NSC were harvested from 11.5 day gestational age (E11.5) pan-EGFP mice (Jackson Laboratory, Bar Harbor, ME), and cultured as described (6 (link)). Fetal intestines were dissected and digested in Collagenase/ Dispase (50 μg/ml; Worthington Biochemical, Freehold, NJ) for 1h. Centrifuged tissue pellets were retrieved and cultured in Dulbecco’s Modified Eagle Medium/Ham’s Nutrient Mixture F12 (DMEM/F12; Corning, Manassas, VA) supplemented 1:1 with N2 supplement (Gibco, Grand Island, NY), with the addition of mouse basic fibroblast growth factor (b-FGF,20 ng/ml; Sigma-Aldrich, St Louis, MO), mouse EGF (20 ng/ml; Sigma-Aldrich, St Louis, MO), 15% chicken embryo extract (Gemini, West Sacramento, CA) and 1% penicillin-streptomycin (Invitrogen, Carlsbad, CA). NSCs proliferated and grew as free-floating neurospheres, and the suspended neurospheres were collected by harvesting the culture medium. Neurospheres were confirmed to contain >95% NSC as determined by staining with the stem cell specific marker Nestin.
+ Open protocol
+ Expand
2

Isolation and Culture of Neural Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
NSC were harvested from 11.5 day gestational age (E11.5) pan-EGFP mice (Jackson Laboratory, Bar Harbor, ME), and cultured as described (6 (link)). Fetal intestines were dissected and digested in Collagenase/ Dispase (50 μg/ml; Worthington Biochemical, Freehold, NJ) for 1h. Centrifuged tissue pellets were retrieved and cultured in Dulbecco’s Modified Eagle Medium/Ham’s Nutrient Mixture F12 (DMEM/F12; Corning, Manassas, VA) supplemented 1:1 with N2 supplement (Gibco, Grand Island, NY), with the addition of mouse basic fibroblast growth factor (b-FGF,20 ng/ml; Sigma-Aldrich, St Louis, MO), mouse EGF (20 ng/ml; Sigma-Aldrich, St Louis, MO), 15% chicken embryo extract (Gemini, West Sacramento, CA) and 1% penicillin-streptomycin (Invitrogen, Carlsbad, CA). NSCs proliferated and grew as free-floating neurospheres, and the suspended neurospheres were collected by harvesting the culture medium. Neurospheres were confirmed to contain >95% NSC as determined by staining with the stem cell specific marker Nestin.
+ Open protocol
+ Expand
3

Porcine Oocyte Maturation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Porcine ovaries were collected from prepubertal gilts at a local slaughterhouse and transported to our laboratory in physiological saline (0.9% NaCl) containing 500 IU/ml of both penicillin and streptomycin at 30-35°C within 2 h kept in a thermos bottle after slaughter. Subsequently ovaries were washed twice with sterile phosphate-buffered saline (PBS), then COCs (cumulus oocyte complexes) were aspirated from antral follicles (2-5 mm in diameter) using a 20-gauge needle attached to a 10-ml disposable syringe. After washing three times with maturation medium, the COCs with intact and compact cumulus were separated from the cellular debris. The medium used for maturation culture was improved TCM-199 supplemented with 75 μg/ml of penicillin, 50 μg/ml of streptomycin, 0.5 μg/ml of LH, 0.5 μg/ml of FSH, 10 ng/ml of epidermal growth factor (mouse EGF; Sigma) and 0.57 mM cysteine (Sigma). To prepare mature oocytes in vitro, a group of 80 COCs were transferred to 500 μl of maturation medium, then medium were subsequently covered with 200 μl paraffin oil in a four-well dish at 38.5°C in a humidified atmosphere of 5% CO2 (Nunc, Roskilde, Denmark).
+ Open protocol
+ Expand
4

Culturing Zebrafish Liver Cell Line ZFL

Check if the same lab product or an alternative is used in the 5 most similar protocols
Zebrafish liver cell line ZFL (Ghosh et al. 1994 (link); Eide et al. 2014 (link)) (CVCL_3276) was cultured in a medium consisting of 50% (v/v) Leibovitz’s L-15 (Sigma-Aldrich, Steinheim, Germany), 35% (v/v) Dulbecco’s modified Eagle's medium (Gibco, Paisley, UK), 15% (v/v) Ham’s Nutrient Mixture F-12 (Gibco, Paisley, UK), and phenol red. Additionally, 150 mg/L sodium bicarbonate (Gibco, Paisley, UK), 15 mM HEPES (Gibco, Paisley, UK), 10 µg/mL bovine insulin (Sigma-Aldrich, Steinheim, Germany), 50 ng/mL mouse EGF (Sigma-Aldrich, Steinheim, Germany), and 5% (v/v) fetal bovine serum (Gibco, Paisley, UK) were supplemented. The cells were cultured in a humidified environment at 28 °C and atmospheric CO2. The cells were passaged weekly in a 1:20 ratio, using PBS (pH 7.4) (Medicago, Uppsala, Sweden) for washing and 0.25% (w/v) trypsin-EDTA (Sigma-Aldrich, Steinheim, Germany) for detachment.
+ Open protocol
+ Expand
5

Spheroid Formation from Murine Bone Marrow and Periodontal Ligament Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells collected by the cell sorter were cultured in non-adherent 24 well plates (Corning, New York, NY, USA) (LepR/Tom+ BM cells: 4781‒9849 cells/well, LepR/Tom BM cells: 18,642‒201,149 cells/well, LepR/Tom+ PDL cells: 83‒348 cells/well, LepR/Tom PDL cells: 9125‒61,003 cells/well) with spheroid-forming media22 (link),27 (link)–29 (link) (1:2 ratio of DMEM/F-12 (1:1) (21331-020) and Human Endothelial Medium (11111-044) supplemented with 3.75% Chicken Extract, 0.1 mM β-ME, 1% Non-essential amino acids, 1% Antibiotic–Antimycotic, 1% N2, 2% B27, 20 ng/mL mouse PDGF-AA (all from Thermo Fisher Scientific), 20 ng/mL human bFGF (ReproCELL Inc., Kanagawa, Japan), 20 ng/mL mouse oncostatin M, 20 ng/mL mouse IGF-1 (all from R&D SYSTEMS), 20 ng/mL mouse EGF (Sigma-Aldrich, St. Louis, MO, USA)). After culturing for 14 days, spheroid-forming efficiency was determined. Fluorescence and phase-contrast images of spheroids were acquired using an All-in-One Fluorescence Microscope (BZ-X700) equipped with a BZ-X-Viewer, BZ-X Analyzer (all from KEYENCE, Osaka, Japan), a CFI Plan Fluor DL (4×/0.13), and a CFI Plan Fluor DL (10×/0.45) (both from Nikon, Tokyo, Japan).
+ Open protocol
+ Expand
6

Cultivation of Zebrafish Liver Cell Line

Check if the same lab product or an alternative is used in the 5 most similar protocols
The zebrafish liver cell line (CVCL_3276) (Ghosh et al. 1994 (link)) was purchased from ATCC (Mannassas, USA). A nutrition medium consisting of 50% (v/v) Leibovitz’s L-15 (Sigma-Aldrich, Steinheim, Germany), 35% (v/v) Dulbecco’s modified Eagle’s medium (Gibco, Paisley, UK), 15% (v/v) Ham’s Nutrient Mixture F-12 (Gibco, Paisley, UK), and phenol red, supplemented by 150 mg/L sodium bicarbonate (Gibco, Paisley, UK), 15 mM HEPES (Gibco, Paisley, UK), 10 μg/mL bovine insulin (Sigma-Aldrich, Steinheim, Germany), 50 ng/mL mouse EGF (Sigma-Aldrich, Steinheim, Germany), and 5% (v/v) fetal bovine serum (Gibco, Paisley, UK), was used for cultivation. The cells were cultured in a humidified environment at 28 °C and atmospheric CO2. Further, cells were passaged weekly in a 1:20 subcultivation ratio. Phosphate buffered saline (PBS, pH 7.4; Medicago, Uppsala, Sweden) was used for washing and 0.25% (w/v) trypsin–EDTA (Sigma-Aldrich, Steinheim, Germany) for cell detachment. All experiments were conducted within passage numbers 5 to 32.
+ Open protocol
+ Expand
7

mCCD Cell Culture Maintenance

Check if the same lab product or an alternative is used in the 5 most similar protocols
mCCDcl1 cells were maintained at 37 °C and 5% CO2 in DMEM/F-12 medium (Thermofisher, 31331093, Waltham, MA, USA), supplemented with 5 µg/mL insulin (Sigma, I-1882, St. Louis, MO, USA), 50 nM dexamethasone (Sigma, D-8893), 60 nM selenium (Sigma, S-9133), 5 µg/mL transferrin (Sigma, T-1428), 1 nM triiodothyronine (Sigma, T-5516), 5 ng/mL mouse EGF (Sigma, E-4127), 100 µg/mL penicillin/streptomycin glutamine (Thermofisher, 10378016), and 2% decomplemented fetal bovine serum (Thermofisher, 10270106). For hypoxic culture, the medium was freshly replaced, and cells were maintained for the indicated time at 0.2% O2 and 5% CO2 in a gas-controlled glove box (InvivO2 400, Baker Ruskinn, Bridgend, UK). Cellular viability was routinely analyzed by trypan blue exclusion using the TC20 automated cell counter (Biorad, Hercules, CA, USA).
+ Open protocol
+ Expand
8

In Vitro Porcine Oocyte Maturation

Check if the same lab product or an alternative is used in the 5 most similar protocols
All protocols performed were approved by the Animal Care and Use Committee of Nanjing Agriculture University and were performed in accordance with Animal Research Institute Committee guidelines. Porcine ovaries were collected from the local slaughterhouse and then transported to laboratory within 3 h in sterile saline (0.9% NaCl) containing 0.03 g/mL of penicillin and 0.03 g/mL of streptomycin at 37°C. Cumulus-oocyte complexes (COCs) were extracted from 3 to 6 mm follicles of ovaries by aspirating with a 20-gauge needle attached to a 5-ml disposable syringe. Oocytes with compact cumulus cells and a uniform ooplasm were selected for in vitro maturation (IVM). The COCs was washed three times with IVM medium [TCM199 (St. Louis, MO, United States,# M2154) supplemented with 75 μg/ml of penicillin, 50 μg/ml of streptomycin, 0.5 μg/ml of LH, 0.5 μg/ml of FSH, 10 ng/ml of epidermal growth factor (mouse EGF, Sigma, #E4127), and 0.57 mM cysteine] and cultured in each well of a four-well dish (Nunc, Roskilde, Denmark) containing 500 μl of IVM medium covered with 200 μl mineral oil at 38.5°C in a humidified atmosphere of 5% CO2 incubator for IVM. After 42–46 h cultivation, COCs were transferred to 0.1% hyaluronidase (w/v) for 3 min at 38.5°C. After three to four rinses with TCM199, matured MII oocytes were collected for next treatment.
+ Open protocol
+ Expand
9

Culturing Zebrafish Liver Cell Line ZFL

Check if the same lab product or an alternative is used in the 5 most similar protocols
Zebrafish liver cell line ZFL26 (link),27 (link) (CVCL_3276) was cultured in a medium consisting of 50% Leibovitz’s L-15 (Sigma-Aldrich, Steinheim, Germany), 35% Dulbecco’s modified Eagle’s medium (Gibco, Paisley, UK), 15% Ham’s Nutrient Mixture F-12 (Gibco, Paisley, UK), and phenol red. Additionally, 150 mg/L sodium bicarbonate (Gibco, Paisley, UK), 15 mM HEPES (Gibco, Paisley, UK), 10 µg/mL bovine insulin (Sigma-Aldrich, Steinheim, Germany), 50 ng/mL mouse EGF (Sigma-Aldrich, Steinheim, Germany), and 5% fetal bovine serum (Gibco, Paisley, UK) were supplemented. The cells were cultured in a humidified environment at 28 °C and atmospheric CO2. The cells were passaged every 3 to 4 days in a 1 to 4 ratio, using PBS (Medicago, Uppsala, Sweden) for washing and 0.25% Trypsin-EDTA (Sigma-Aldrich, Steinheim, Germany) for detachment.
+ Open protocol
+ Expand
10

Expansion and Maintenance of Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols
HEK293T cells (ATCC, CRL-3216) and human newborn foreskin fibroblasts (HFFs, from ATCC, SCRC-1041) were purchased from ATCC. The SCR029 cells (Chemicon, SCR029) are well-characterized mouse cortical NSCs obtained from Chemicon. These cells were tested for mycoplasma contamination before experiments and they are negative. HEK293T cells, HFFs, and MEFs were all expanded in the MEF medium [Dulbecco’s modified Eagle’s medium (DMEM, Hyclone), 10% fetal bovine serum (FBS) (Gibco), 1 × Pen/Strep (Gibco), 1 × MEM non-essential amino acids (Gibco), and 0.008% (v/v) 2-mercaptoethanol (Sigma)]. All the NSCs were plated on culture dishes pre-coated with 5 μg/ml poly-l-ornithine (Sigma) and 5 μg/ml laminin (Sigma). miNSCs and human iNSCs were cultured in the NSC medium containing the N3 medium supplemented with 10 ng/ml recombinant mouse EGF (R&D systems) and 10 ng/ml recombinant human bFGF (R&D Systems), with or without 2 ng/ml Dox (Sigma). The N3 medium contains DMEM/F12 (Life Technologies) with 1 × Pen/Strep, 25 μg/ml insulin (Sigma), 50 μg/ml Apo-transferrin (Sigma), 1.28 ng/ml progesterone (Sigma), 16 ng/ml putrescine (Sigma), and 0.52 μg/ml sodium selenite (Sigma). The SCR029 cells were cultured in the Neural Stem Cell Expansion Medium (Chemicon, SCM003) supplemented with 20 ng/ml mouse EGF, 20 ng/ml human bFGF, and 2 μg/ml heparin (Sigma).
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!