MenSC and umbilical cord (UC) MSC were isolated from menstrual fluid of the umbilical cord of three different healthy donors each and characterized according to ISCT31 (link) criteria as we have previously described15 (link),22 (link),32 . Human dermal fibroblasts were purchased from Lonza (cat. CC-2511, Lonza, Walkersville, MD USA). Human umbilical cord vein endothelial cells (HUVEC) and human microvascular endothelial cell line (HMEC-1) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). HUVEC and fibroblasts were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (15-018-CV, Corning, New York, USA) supplemented with 10% fetal bovine serum (FBS) (Lonza, Walkersville, MD USA), 1% Penicillin/Streptomycin (P/S) (Life Technologies, Carlsbad, CA, USA) and 1 mM L-glutamine (Life Technologies). HMEC-1 cells were cultured in endothelial cells growth media 2 (EGM-2) (cc4147, Lonza). All cells were regularly tested for mycoplasma contamination using a mycoplasma detection kit (G238, Applied Biological Materials Inc., Richmond, BC, Canada). All cells were cultured in humidified incubation chambers at 37 °C with 5% CO2.
Hmec 1
Manufactured by Lonza
Sourced in Switzerland
The HMEC-1 is a laboratory cell line derived from human mammary epithelial cells. It is a well-characterized in vitro model that can be used for various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Hmec 1, by American Type Culture Collection (2 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Lonza (1 mentions)
Human umbilical vein endothelial cells (huvec), by Corning (1 mentions)
Egm 2, by Lonza (1 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions)
Il 1a, by R&D Systems (1 mentions)
Huvecs, by Lonza (1 mentions)
Hmec 1, by iCell Bioscience (1 mentions)
Ebm 2, by Lonza (1 mentions)
Egm 2mv medium, by Lonza (1 mentions)
Normal human dermal fibroblasts (nhdf), by Lonza (1 mentions)
Normal human dermal fibroblasts (nhdf), by American Type Culture Collection (1 mentions)
5 protocols using hmec 1
1
Isolation and Characterization of Mesenchymal Stem Cells
2
Exosome Modulation of Cytokine Release
3
Overexpression of Codon-Optimized ELTD1 in Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human umbilical cord endothelial cells (HUVECs) and human microvascular cells (HMEC-1) were purchased from Lonza (Basel, Switzerland) and cultured in their EGM2 and EGM2-MV media respectively. IL8 and IL1a (R&D Systems, Minneapolis, MN, USA) were added to cells at 10 ng/mL. IL1-receptor antagonist (Sigma, St. Louis, MO, USA) was added at 1 μg/mL. The endogenous sequence of ELTD1 is poorly expressed therefore the cDNA was codon optimised using the JCat bioinformatics tool (http://www.jcat.de/ ) (accessed on 31 November 2017) without altering the amino acid coding sequence. Codon optimised ELTD1 (coELTD1) and HA-tagged coELTD1 were cloned into pLenti6.2V5DEST (ThermoFisher, Waltham, MA, USA) and the vector alone was used as a control. Virus was produced in 293T and concentrated by ultracentrifugation using standard techniques. The viruses were titred using blasticidin resistance and endothelial cells infected at MOI 5 for all experiments.
4
Modeling Endothelial Dysfunction in Diabetes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human microvascular endothelial cells (HMEC-1) (iCell Bioscience Inc., Shanghai, China) were cultured in EGM-2MV medium (Lonza, Basel, Switzerland) supplemented with fetal bovine serum (FBS) and growth factors at 37 °C in the presence of 5% CO2 in the air. To mimic diabetes in vitro, HMEC-1 cells were subjected to endothelial basal medium (EBM-2, Lonza) containing 25 mmol/L d -glucose (Aladdin, Shanghai, China) (high glucose, HG). To simulate ischemia-induced tissue starvation, cells were cultured in medium supplemented with no additional growth factors (nutrient deprivation, ND) (Caporali et al. 2011 (link)). Cells incubated with d -Mannitol (normal glucose, NG) and EGM-2MV medium supplemented with FBS and growth factors served as controls.
Prior to cell seeding, cells were infected with prepared Ad-AURKAOE or Ad-NCOE at a multiplicity of infection of 10 for 48 h. Cells were then detached and re-seeded on plates for subsequent analyses under different stimuli as described above.
Prior to cell seeding, cells were infected with prepared Ad-AURKAOE or Ad-NCOE at a multiplicity of infection of 10 for 48 h. Cells were then detached and re-seeded on plates for subsequent analyses under different stimuli as described above.
5
Comparative Cell Culture Protocol for Glioblastoma and Normal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
U-87 MG glioblastoma cell line (cancerous),
HMEC-1 (noncancerous endothelial cells), and NHDF (noncancerous fibroblasts)
were purchased from the American Type Culture Collection (ATCC) and
Lonza, respectively. U-87 MG cells were cultured in EMEM with 10%
FBS, 2 mM glutamine, and 1% (100 U/mL) penicillin–streptomycin;
HMEC-1 cells were cultured in MCDB 131 with 10% heat-inactivated FBS,
2 mM glutamine, 1% penicillin–streptomycin, and 10 ng/mL epidermal
growth factor (EGF human protein) and NHDF in fibroblast growth medium
supplemented with fibroblast growth kit (Lonza), respectively. U-251
MG glioblastoma cells transfected with dsRed66 (link) were kindly provided by Manon Carré and cultured in DMEM
with phenol red, 2 mM glutamine, 10% FBS, and 1% penicillin–streptomycin.
All cells were routinely maintained at 37 °C and 5% CO2.
HMEC-1 (noncancerous endothelial cells), and NHDF (noncancerous fibroblasts)
were purchased from the American Type Culture Collection (ATCC) and
Lonza, respectively. U-87 MG cells were cultured in EMEM with 10%
FBS, 2 mM glutamine, and 1% (100 U/mL) penicillin–streptomycin;
HMEC-1 cells were cultured in MCDB 131 with 10% heat-inactivated FBS,
2 mM glutamine, 1% penicillin–streptomycin, and 10 ng/mL epidermal
growth factor (EGF human protein) and NHDF in fibroblast growth medium
supplemented with fibroblast growth kit (Lonza), respectively. U-251
MG glioblastoma cells transfected with dsRed66 (link) were kindly provided by Manon Carré and cultured in DMEM
with phenol red, 2 mM glutamine, 10% FBS, and 1% penicillin–streptomycin.
All cells were routinely maintained at 37 °C and 5% CO2.
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!