Hrmecs

Manufactured by Merck Group

Sourced in United States

HRMECs are primary human renal microvascular endothelial cells. They are isolated from human kidney tissue and cultured in vitro. HRMECs are used for research purposes to study vascular biology, angiogenesis, and renal function.

Automatically generated - may contain errors

Lab products found in correlation

5 protocols using hrmecs

Retinal Microvascular Endothelial Cell Proliferation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human Retinal Microvascular Endothelial cells (HRMECs) were purchased from Innoprot. Cells were derived from adult eyes of male donors. HRMECs were cultured on flasks coated with fibronectin from human plasma (F2006, Sigma-Aldrich) and in Endothelial Cell Medium (ECM, Innoprot) composed of 5% Fetal Bovine Serum (FBS), Endothelial Cell Growth Supplements (ECGS) and penicillin/streptomycin. HRMECs were grown in normal (5 mM D-glucose) and high-glucose (25 mM D-Glucose) conditions. Cells were cultured in 5 mM D-glucose plus 20 mM l-Glucose as the osmotic control. HRMECs between passage 4 (P4) and P5 were used as early passage (ep) controls. Cells were treated with high-glucose conditions for 4 weeks. RNA and protein were sampled after 4-week-culture under high glucose conditions when HRMECs were P11-P16 and defined as late passage (lp). Hayflick limit was reached at ≥ P20. To define HRMEC proliferation capacity in vitro, growth curves were generated. Cells were passaged every 3–5 days and seeded at a known density (2 × 10⁴/ml) to measure population doubling level (PDL) at each passage, until they reached Hayflick limit. The CASY cell counter analyzer (Cambridge Bioscience) was used for cell counts at every passage.

Bertelli P.M., Pedrini E., Hughes D., McDonnell S., Pathak V., Peixoto E., Guduric-Fuchs J., Stitt A.W, & Medina R.J. (2022). Long term high glucose exposure induces premature senescence in retinal endothelial cells. Frontiers in Physiology, 13, 929118.

+ Open protocol

+ Expand

HEK293 and HRMEC Protein Extraction Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEK293 cells were grown in Dulbecco’s modified Eagles medium (DMEM; Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum in humidified 5% CO₂ at 37°C. Wild-type (WT) or mutant pCDNA3.1-Sirt6-Flag plasmids were transfected into HEK293 cells using lipofectamine 2000 and Opti-MEM I reagents (Life Technologies) according to the manufacturer’s instructions. 48 hours after transfection, cells were collected and homogenized in ice-cold RIPA Lysis and Extraction Buffer (Thermo Fisher Scientific, Rockford, IL). The homogenates were centrifuged at 13800 g for 10 minutes at 4°C. Supernatants were collected, and protein concentrations were determined by a bicinchoninic acid assay (Thermo Fisher Scientific). Samples were stored at -80°C until used.
Primary human retinal microvascular endothelial cells (HRMECs) were purchased from Cell Systems (Kirkland, WA) and cultured in medium constituted of 50% CSC Serum-Containing Medium (Cell Systems) and 50% EGM^™ Endothelial Cell Medium (Lonza Walkersville Inc., Walkersville, MD). HRMECs were permeabilized with 40 μg/ml digitonin (Sigma-Aldrich) for 5 minutes, then treated with 5 mM SIN-1 for 2 hours in Endothelial Basal Medium (Lonza Walkersville Inc.) at 37°C with 5% CO₂ before cells were collected and homogenized in ice-cold RIPA Lysis and Extraction Buffer [19 (link)].

Hu S., Liu H., Ha Y., Luo X., Motamedi M., Gupta M.P., Ma J.X., Tilton R.G, & Zhang W. (2014). Posttranslational Modification of Sirt6 Activity by Peroxynitrite. Free radical biology & medicine, 79, 176-185.

+ Open protocol

+ Expand

Culturing Human Retinal Microvascular Endothelial Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

HRMECs were purchased from American type culture collection (ATCC). HRMECs were cultured in Dulbecco's modified eagle's medium (DMEM) (Sigma-Aldrich, USA) supplemented with 1% penicillin/streptomycin (100 mg/L, Gibco, USA) and 10% heat-inactivated fetal bovine serum (FBS) (Gibco, USA) at 37 °C in 5% CO₂ atmosphere. For normal glucose and high glucose conditions, 5 mM and 33 mM D-glucose (Gibco, USA) were added to the medium for 48 h, respectively.

Xu Y., Zou H., Ding Q., Zou Y., Tang C., Lu Y, & Xu X. (2022). tiRNA-Val promotes angiogenesis via Sirt1–Hif-1α axis in mice with diabetic retinopathy. Biological Research, 55, 14.

+ Open protocol

+ Expand

High Glucose-induced HMGB1 Regulation in HRMECs

Check if the same lab product or an alternative is used in the 5 most similar protocols

HRMECs were purchased from Procell Life Science&Technology (Wuhan, China). The HRMECs were grown on the poly‐L‐lysine‐coated plates in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 5% fetal bovine serum (FBS), 100 units/mL penicillin, 100 μg/mL streptomycin, endothelial cell growth factor, heparin, and hydrocortisone in a 5% CO₂‐enriched atmosphere with constant humidity. To maintain uniform conditions, all experiments were performed using passage 5 HRMECs. HRMECs were grown in the 50 mM glucose medium (Sigma‐Aldrich) for 48 h. D‐Mannitol was used as an osmotic control. For the treatment, NK (1 μM and 2 μM) was pretreated for 2 h before HG stimulation. For HMGB1 silencing, siRNA constructs were diluted in buffer (Lipofectamine transfection reagent; Invitrogen Inc., Carlsbad, CA, USA) and transfected in culture medium of HRMECs at a working concentration of 100 nM for 24 h before NK treatment. Scrambled siRNA was used as control. The sequences for HMGB1‐siRNA were 5′‐GGC UUU CAC UUA AGA ACU UTT‐3′.

Huang Z., Chu W.K., Ng T.K., Chen S., Liang J., Chen C., Xu Y., Xie B., Ke S., Liu Q., Chen W, & Huang D. (2023). Protective effects of nattokinase against microvasculopathy and neuroinflammation in diabetic retinopathy. Journal of Diabetes, 15(10), 866-880.

+ Open protocol

+ Expand

HRMECs Glucose Metabolism Regulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

The HRMECs were purchased from Cell Systems, and were routinely cultured in M199 medium (EMD Millipore) supplemented with 100 units of penicillin and 100 µg of streptomycin (both from Sigma-Aldrich; Merck KGaA) per milliliter of medium. All cells (passages 5-12) were cultured in grade plastic-ware and maintained in an atmosphere of 5% CO₂ at 37°C.
For the establishment of the HG cell models, the HRMECs were cultured in conditioned medium with 5 mM [serving as the normal glucose (NG) group] or 30 mM (the HG group) D-glucose (Sigma-Aldrich; Merck KGaA) and incubated at 37°C with 5% CO₂; the HG group was then treated with or without 1 µM of the NLRP3 inhibitor, CY-09, 1 µM of the ASK1 inhibitor, GS-444217 or 10 µM of the p38 inhibitor, SB203580 (all from MedChemExpress) for 24 h, respectively. Each inhibitor was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 50 mM for use as stock solutions that were diluted to the required concentrations for in vitro experiments.

Zou W., Luo S., Zhang Z., Cheng L., Huang X., Ding N., Pan Y, & Wu Z. (2020). ASK1/p38-mediated NLRP3 inflammasome signaling pathway contributes to aberrant retinal angiogenesis in diabetic retinopathy. International Journal of Molecular Medicine, 47(2), 732-740.

+ 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?

Revolutionizing how scientists
search and build protocols!