HMECs were purchased from Lonza (Walkersville, MD, USA) and cultured in MCDB170 (US Biological, Salem, MA, USA) supplemented with 0.1 mM ethanolamine (Sigma-Aldrich, St Louis, MO, USA), 0.1 mM ortho-phosphoethanolamine (Sigma), 0.25% bovine pituitary extract (Hammond Cell Tech, Windsor, CA, USA), 5 ng/ml EGF (Peprotech, Rocky Hill, NJ, USA), 0.5 μg/ml hydrocortisone (Sigma), 5 μg/ml insulin (Sigma), 5 μg/ml transferrin (Sigma), and 5 μM isoproterenol (Sigma).44 (link) For immortalization, HMECs were transduced with hTERT-expressing retrovirus as described previously.17 (link)For suspension culture, cells were detached by treatment with 0.025% trypsin, followed by resuspension in the growth medium supplemented with 0.5% methylcellulose and plated on polyHEMA-coated dishes. Anisomycin, pepstatin A, and polyHEMA were obtained from Sigma-Aldrich. E64d and Z-VAD-fmk (Z-VAD) were purchased from Peptide Institute, Inc. (Osaka, Japan), staurosporine (STS) and bafilomycin from Wako Pure Chemical Industries, Ltd. (Osaka, Japan), and necrostatin-1 from Enzo Life Sciences, Inc. (Farmingdale, NY, USA). BD Matrigel Matrix Growth Factor Reduced (GFR) was obtained from BD Biosciences (Franklin Lakes, NJ, USA).
Matrigel matrix growth factor reduced gfr
Manufactured by BD
Sourced in United States
BD Matrigel Matrix Growth Factor Reduced (GFR) is a soluble basement membrane extract of proteins derived from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. It is used as a 3D cell culture matrix to support the growth and differentiation of various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Hmecs, by Lonza (1 mentions)
Hmecs, by Merck Group (1 mentions)
Staurosporine sts, by Fujifilm (1 mentions)
Pepstatin a, by Merck Group (1 mentions)
Poly hema, by Merck Group (1 mentions)
Necrostatin 1, by Enzo Life Sciences (1 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Insulin, by Merck Group (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Ethanolamine, by Merck Group (1 mentions)
Anisomycin, by Merck Group (1 mentions)
Transferrin, by Merck Group (1 mentions)
Isoproterenol, by Merck Group (1 mentions)
Mcdb170, by US Biological (1 mentions)
Z vad fmk, by Peptide Institute (1 mentions)
Axio observer microscope, by Zeiss (1 mentions)
Stempro hesc, by Thermo Fisher Scientific (1 mentions)
Liberase dh, by Roche (1 mentions)
3 protocols using matrigel matrix growth factor reduced gfr
1
Immortalized Human Mammary Epithelial Cell Culture
2
Capillary Tube Formation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BD Matrigel Matrix Growth Factor Reduced (GFR) (BD Biosciences, San Jose, CA, USA) was used to promote the differentiation of HRMECs into capillary tube–like structures. A total of 100 µL of thawed BD Matrigel Matrix GFR was added to 96-well tissue culture plates, followed by incubation at 37°C for 60 minutes to allow polymerization. Prior to the Matrigel assays, HRMECs were cultured in EGM-2 Medium to 70% confluence in 100-mm tissue culture dishes. On the day of the assay, HRMECs were harvested and resuspended in EGM-R Medium. Subsequently, HRMECs (2.0 × 105 cells/mL) were seeded on the polymerized BD Matrigel Matrix GFR in EGM-R Medium, supplemented with the indicated concentrations of pNaKtide, and incubated for 24 hours at 37°C. Tubes were observed using a ZEISS Axio Observer microscope (Carl Zeiss Microscopy, White Plains, NY, USA) at 5× magnification. At least three independent fields per sample were captured. Tube networks were measured by a blinded investigator to determine the total tube length and total tube numbers per field using ImageJ (National Institutes of Health, Bethesda, MD, USA) with the Angiogenesis Analyzer plugin, as described previously.39 (link),40 The data were normalized to controls. Each sample was tested in triplicate, and the entire experiment was performed three separate times.
3
Primary Colorectal Cancer Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Preparation and culture of primary colorectal cancer cells were performed using the CTOS method [28] (link). Briefly, surgical samples or xenograft tumors of NOD/SCID mice were partially digested with Liberase DH (Roche, Mannheim, Germany) and filtered through cell strainers. Fragments on the 100-µm or 40-µm cell strainer (BD Falcon, Franklin Lakes, NJ) were collected and cultured in StemPro hESC (GIBCO). For hypoxic culture, CTOS samples were embedded in BD Matrigel Matrix Growth Factor Reduced (GFR) (BD Biosciences, San Jose, CA) and cultured in StemPro hESC or basal medium (DMEM F12/Glutamax, 0.1 mM 2-mercapthoethanol, 2% bovine serum albumin). For the chemosensitivity assay during the inactive period, CTOS samples were pre-cultured in basal medium under 1% O2 conditions for 7 days, and then treated with 5-FU or SN38. After 7 days of exposure to these drugs, the CTOS samples were re-oxygenated, and the culture medium was changed to fresh StemPro hESC.
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!