Human retinal microvascular ECs (HRMECs) (Angio-Proteomie, Boston, MA, USA) were cultured in endothelial growth medium (EGM) (Angio-Proteomie, USA) containing 10% fetal bovine serum (FBS), recombinant growth factors, and 1× penicillin and streptomycin. Cells were grown on Quick Coat Solution (Angio-Proteomie, USA) coated plates and maintained in humidified 5% CO2 /95% air at 37 °C.
Hrmecs
Manufactured by Angio-Proteomie
Sourced in United States
HRMECs are primary human retinal microvascular endothelial cells. They are an in vitro cell model derived from human retinal tissue.
Automatically generated - may contain errors
Lab products found in correlation
Hrmecs, by Thermo Fisher Scientific (2 mentions)
Quick coat solution, by Angio-Proteomie (1 mentions)
Withaferin a, by ChromaDex (1 mentions)
Hek293 cell, by American Type Culture Collection (1 mentions)
Newborn calf serum ncs, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Mir nc mimic, by RiboBio (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Lentivirus, by Geneseed (1 mentions)
Mir 20b 5p inhibitor, by RiboBio (1 mentions)
Penicillin streptomycin solution, by ScienCell (1 mentions)
Crl 2647, by American Type Culture Collection (1 mentions)
Endothelial cell medium mv, by PromoCell (1 mentions)
Endothelial cell supplement mix, by PromoCell (1 mentions)
Crl 2648, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by ScienCell (1 mentions)
8 protocols using hrmecs
1
Culturing Human Retinal Microvascular Endothelial Cells
2
In Vitro Model of Retinal I/R Injury
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary human retinal microvascular endothelial cells (HRMECs, Cat# cAP-0010, Angio Proteomie, Boston, MA, USA), passages 2–5, were cultured in endothelial basal media containing 5% fetal bovine serum and premixed endothelial cell growth supplement. Upon reaching 100% confluence, cells were serum starved for 3 h and then treated with 1% fetal bovine serum.
To simulate retinal I/R injury at the cellular level, HRMECs were cultured in plates and subjected to hypoxia (1% O2) for 3, 6, 12, or 24 h in D-Hanks buffer followed by 2 h of reoxygenation (5% O2) in complete DMEM. To further study the impact of I/R-induced oxidative stress, cultured HRMECs were subjected to 2 h of hydrogen peroxide (H2O2) at different concentrations (50, 100, 200, 400, or 800 µmol/l). At the same time, the effects of Withaferin A were tested. Withaferin A was purchased from ChromaDex (Cat# 5119-48-2, Irvine, CA, USA).
To simulate retinal I/R injury at the cellular level, HRMECs were cultured in plates and subjected to hypoxia (1% O2) for 3, 6, 12, or 24 h in D-Hanks buffer followed by 2 h of reoxygenation (5% O2) in complete DMEM. To further study the impact of I/R-induced oxidative stress, cultured HRMECs were subjected to 2 h of hydrogen peroxide (H2O2) at different concentrations (50, 100, 200, 400, or 800 µmol/l). At the same time, the effects of Withaferin A were tested. Withaferin A was purchased from ChromaDex (Cat# 5119-48-2, Irvine, CA, USA).
3
Effects of CTRP3 on Retinal Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human retinal microvascular endothelial cells (HRMECs) (Cat# cAP-0010, Angio-Proteomie, Boston, MA, USA) were plated on six-well plates and cultured at 37 °C in a 5% CO2 incubator. Upon reaching 100% confluence, HRMECs were randomized to receive one of the following treatments: (1) normal glucose normal lipids (NGNL, 5.5 mM D-glucose/19.5 mM L-glucose,) receiving vehicle or gCTRP3 (0.3, 1, and 3 μg/mL, Cat# 00082-01; Aviscera Bioscience, Santa Clara, CA, USA) for 24 h; (2) high glucose/high lipids (HGHL, 25 mM D-glucose/250 μM palmitates) [19 (link)], receiving vehicle or gCTRP3 treatment (0.3, 1, and 3 μg/mL) for 24 h; 25 mM L-glucose plus fatty acid-free BSA was used as an osmotic control [20 (link)].
4
Culturing Endothelial and HEK293 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HRMECs were obtained from Angio-Proteomie (Peabody, MA, United States). HRMECs were cultured in endothelial cell medium (ECM) supplemented with 5% Newborn Calf Serum (NCS; Life Technologies), 20 μg/ml of ECGS, 100 μg/ml streptomycin, and 100 U/ml penicillin in a T-25 flask coated with 8.5 μg/ml of BPF at 37°C with 5% CO2. When the cells became 90% confluent, they were subcultured at a 1:3 ratio. HEK293 cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, United States) and cultured in DMEM supplemented with 10% NCS and incubated at 37°C with 5% CO2.
5
Investigating miR-20b-5p and circDNMT3B in HRMECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HRMECs (Angio-Proteomie, Boston, Mass, USA) were cultured in endothelial cell medium (ECM) with 5% fetal bovine serum, 1% endothelial cell growth supplement, and 1% penicillin/streptomycin solution (ScienCell Research Laboratories, Carlsbad, CA, USA) at 37 °C under 5% CO2 atmosphere. Cells between passages three and eight were used in this study. Cells were treated with 5 mM glucose as a normal glucose (NG) control, 5 mM glucose plus 25 mM mannitol as an osmotic control, or 30 mM glucose as a high-glucose (HG) treatment. miR-20b-5p inhibitor, miR inhibitor negative control (NC), miR-20b-5p mimic, or miR mimic NC (RiboBio, Guangzhou, China) was transfected into HRMECs using Lipofectamine 3000 (Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions. Small interfering BMP and activin membrane-bound inhibitor (siBAMBI) and siRNA NC synthesized by RiboBio were transfected using Lipofectamine RNAiMAX (Life Technologies). The human circDNMT3B sequence was inserted into the pLCDH-ciR vector, and then incorporated into a lentivirus by Geneseed Biotech (Guangzhou, China). To induce stable circDNMT3B-overexpressing HRMECs, the HRMECs at passage three were infected with lentivirus containing circDNMT3B or ciR NC (empty vector). All sequences used in this study are listed in Supplementary Table S1.
6
Wnt-3a Conditional Medium Preparation
7
Primary Cell Culture and Animal Use
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HRMECs (Angio-Proteomie, Boston, MA) are primary cells that were used for in vitro studies and were cultured as previously described [17 (link)]. Neonatal mice (C57BL/6J) and adult mice (C57BL/6J, 20~25 g) were purchased from the animal center of Inner Mongolia University and were raised in the animal room of the Affiliated Hospital of Inner Mongolia University for the Nationalities. This study adhered to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and was performed in accordance with the guidelines provided by the Animal Care and Use Committee of the Affiliated Hospital of Inner Mongolia University for the Nationalities. The animals were housed with free access to laboratory food and water under a 12:12 h light:dark cycle.
8
Culturing Primary HRMECs with Glucose
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell culture Primary HRMECs were purchased from Angio-Proteomie (Oxfordshire, UK) and cultured in endothelial cell medium supplemented with 5% fetal bovine serum (ScienCell, Carlsbad, CA). Cells from passage 3 to 10 were used and incubated with normal D-glucose (5.5 mM), high D-glucose (25 mM) and L-glucose (25 mM, as osmotic control).
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!