All the chemicals used in this study are commercially purchased and used without further purification. Lipofectamine (LPS) was purchased from Thermo Fisher (00‐4976‐93); Beta Amyloid (1–42) was procured from Anaspec (AS24224), Mammalian protein extraction reagent (MPER; #78 505)), EDTA, Protease inhibitor (78 425) were purchased from Thermo Fischer Scientific. For 2D cell culture, murine microglia BV2 cell was used, human microglia HMC3 and cultured in Dulbecco's modified Eagle's media (12‐707F; Lonza) supplemented with 10% fetal bovine serum (FBS), L‐Glutamine (25‐005‐Cl; Corning), and Penicillin/streptomycin (DE17‐602E; Lonza). Cells were cultured in a 37 °C incubator with water‐saturated air and a 5% CO2 atmosphere and splitting was done every 2–3 days using trypsin (BE17‐161E; Lonza).
Be17 161e
Manufactured by Lonza
The BE17-161E is a laboratory instrument manufactured by Lonza. It is designed for use in scientific research and analysis applications. The core function of this equipment is to perform specific tasks within a controlled laboratory environment. No further details on the intended use or capabilities of this product can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Protease inhibitor, by Thermo Fisher Scientific (1 mentions)
Mammalian protein extraction reagent, by Thermo Fisher Scientific (1 mentions)
De17 602e, by Lonza (1 mentions)
Dulbecco s modified eagle s media, by Lonza (1 mentions)
Beta amyloid 1 42, by AnaSpec (1 mentions)
Penicillin streptomycin, by Lonza (1 mentions)
L glutamine, by Corning (1 mentions)
Trypsin, by Lonza (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Supplementmix, by PromoCell (1 mentions)
2 protocols using be17 161e
1
Microglia Cell Culture Protocols
2
HUVEC Isolation from Umbilical Cords
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Primary human umbilical vein endothelial cells (HUVECs) were isolated from human umbilical cords obtained at the Leiden University Medical Centre after written informed consent was obtained and the umbilical cord was collected and processed anonymously. The umbilical cord was rinsed with phosphate-buffered saline (PBS) to remove any remaining blood. To detach the endothelial cells from the umbilical cord, trypsin/EDTA (BE17-161E; Lonza) was used. After 20 min of incubation at 37 C, the trypsin was inactivated with 20% fetal bovine serum in PBS, and the entire solution in the umbilical cord was collected. The umbilical vein was flushed with PBS one more time to ensure that all detached cells were collected before centrifugation at 300 g for 7 min. Cells were then cultured in 1% gelatin pre-coated flasks in endothelial growth medium (EGM2 medium, C-22011 supplemented with SupplementMix; Promocell) with 1% antibiotics (penicillin/streptomycin, 15 070 063; Gibco). We cultured HUVECs from two different donors (one boy and one girl). We pooled them together and froze them in several vials for future experiments once they were confluent.
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