Human cerebral microvascular endothelial cells (HBEC-5i, ATCC® CRL-3245) and Human-breast cancer cell line MDA-MB-231 (ATCC® HTB-26) were purchased from American Type Culture Collection (ATCC, United States). HBEC-5i cells were cultured as a monolayer on 0.1% gelatin solution (Gibco/Thermo Fisher, United States) coated T-flasks in DMEM:F12 medium (Gibco/Thermo Fisher, United States) supplemented with 10% FBS (Gibco/Thermo Fisher, United States), 1% penicillin/streptomycin antibiotic solution (Gibco/Thermo Fisher, United States), and 40 μg/mL endothelial growth supplement (ECGS) (Sigma-Aldrich, Spain), according to the manufacturer’s instructions. MDA-MB-231 cells were cultured as a monolayer in DMEM medium (Gibco/Thermo Fisher, United States) supplemented with 10% FBS and 1% penicillin/streptomycin antibiotic solution, according to manufacturer’s instructions. Both cells were grown in a humidified atmosphere of 5% CO2 at 37°C (MCO-18AIC (UV), Sanyo, Japan) with the medium changed every other day.
Gelatin solution
Manufactured by Thermo Fisher Scientific
Sourced in United States
Gelatin solution is a laboratory product that serves as a stabilizing agent and gelling compound. It is a clear, viscous liquid derived from the partial hydrolysis of collagen. Gelatin solution is commonly used in various life science applications to provide structural support and enhance the consistency of samples or solutions.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Mda mb 231, by Thermo Fisher Scientific (1 mentions)
Atcc htb 26, by American Type Culture Collection (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Hbec 5i, by American Type Culture Collection (1 mentions)
Penicillin streptomycin antibiotic solution, by Thermo Fisher Scientific (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Hbec 5i, by Thermo Fisher Scientific (1 mentions)
Mco 18aic, by Sanyo (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
Nci h1975, by American Type Culture Collection (1 mentions)
Nci h1975, by Thermo Fisher Scientific (1 mentions)
Hff 1, by American Type Culture Collection (1 mentions)
Collagenase 2, by Worthington (1 mentions)
Amphotericin b, by Thermo Fisher Scientific (1 mentions)
Amphotericin b, by Merck Group (1 mentions)
3 protocols using gelatin solution
1
Culturing Human Endothelial and Breast Cancer Cells
2
Cultivation of NSCLC and Cardiomyocyte Cells
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NSCLC cell lines NCI-H1975 and A549 and human foreskin fibroblasts (HFF-1) were purchased from ATCC. NCI-H1975 and A549 cells were cultured in RPMI-1640 (Life Technologies) supplemented with 10% fetal bovine serum (FBS; Corning) and 1× penicillin‒streptomycin (Gibco). HFF-1 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS (Corning) and 1× penicillin‒streptomycin (Gibco). Primary cardiomyocytes were isolated from neonatal Sprague‒Dawley rats (Zhejiang Academy of Medical Sciences). Ventricles from the heart were sliced into approximately 1 mm sections in Hanks balanced salt solution (HBSS; Gibco). After digestion with collagenase II (0.2 mg/ml; Worthington) and trypsin (0.3 mg/ml, Gibco) for 1 h at 37 °C, cardiomyocytes were collected by centrifugation. Cells were then transferred to a 96-well plate or the biosensor chip previously coated with gelatin solution (Gibco). DMEM supplemented with 10% FBS and 1× penicillin‒streptomycin was used to maintain cardiomyocytes.
3
Isolation and Expansion of Renal Epithelial Cells from Urine
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Putative renal epithelial cells were collected and expanded from urine with minor modifications from methods previously described 11 , 12 . Briefly, urine samples were collected from 10 individuals with DS and one individual without DS and centrifuged at 400g for 10 minutes. The pellet was washed in buffer containing 1X Dulbecco's phosphate‐buffered saline (DPBS) with Penicillin/Streptomycin (PS, Hyclone, Pittsburgh, PA, www.gelifesciences.com ), 0.5 µg/ml Amphotericin B (Sigma‐Aldrich, St. Louis, MO, www.sigmaaldrich.com ). The pellet was resuspended with primary culture media containing Renal Cell Growth Medium (REGM, Lonza, Basel, Switzerland, www.lonza.com ), 10% FBS (Gibco, Waltham, MA, www.thermofisher.com ), PS, and 0.5 µg/ml Amphotericin B. Cells were then transferred to 12‐well tissue culture dishes coated with 1% gelatin solution (Gibco). During the first 3 days, 1 ml of primary culture medium was added. Starting on the fourth day, most of the medium was aspirated and 1.5 ml of REGM was added. Then, this procedure was repeated every other day until the T21 urine‐derived cells had expanded enough to cover above 70% of the plate.
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