Fibroblasts were derived from colorectal cancer patients. NFs were isolated from normal colon segment with a distance of approximately 20 cm from the tumor and TAFs were isolated from the tumor of the same patient with moderately differentiated adenocarcinoma and lymph node metastasis. Tissue pieces were washed rigorously with phosphate buffered saline (PBS), antibiotic-antimycotic (10×) (Lonza), gentamycin (10×) (Lonza) and cut into pieces for incubation overnight at 37 °C in DMEM (Euroclone, Milano, IT), antibiotic-antimycotic (1×), gentamycin (1×) in the presence of collagenase type II 150 U/ml (Gibco). After incubation, the solution was centrifuged briefly to remove undigested tissue pieces and plated on gelatin (1 g/l) pre-treated dishes in αMEM (Euroclone) supplemented with 10% defined FBS (GE-Healthcare), non-essential amino acids (Euroclone), L-alanine-l -glutamine (Euroclone), penicillin/streptomycin (5×) (Euroclone), antibiotic-antimycotic (5×), and gentamycin (5×).
L alanine l glutamine
Manufactured by Euroclone
Sourced in United States
L-alanine-l-glutamine is a laboratory product that serves as a chemical compound. It consists of the amino acids L-alanine and L-glutamine.
Automatically generated - may contain errors
Lab products found in correlation
Collagenase type 2, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Euroclone (2 mentions)
Gentamycin, by Thermo Fisher Scientific (2 mentions)
Non essential amino acids (neaa), by Euroclone (2 mentions)
Gentamycin, by Euroclone (2 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (2 mentions)
Antibiotic antimycotic, by Euroclone (2 mentions)
Antibiotic antimycotic, by Lonza (2 mentions)
Gentamycin, by Lonza (2 mentions)
Dulbecco modified eagle medium (dmem), by Euroclone (1 mentions)
2 protocols using l alanine l glutamine
1
Fibroblast Isolation from Colorectal Cancer
2
Isolation of Colon Fibroblasts from Tumor and Normal Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Normal colon naive fibroblasts and tumour-associated fibroblasts (TAFs) were derived from the same patient diagnosed with colon adenocarcinoma. The naive fibroblasts were isolated from the normal colon segment approximately 20 cm from the tumour close to the edge of the isolated colon segment that was histologically free of neoplasia. The isolated tissues were washed rigorously in phosphate-buffered saline (PBS), an antibiotic-antimycotic (Lonza, Basel, Switzerland), gentamycin (Lonza) and cut into pieces for incubation overnight at 37 °C in αMEM (Euroclone, Milano, IT) with antibiotic-antimycotic and gentamycin in the presence of collagenase type II 150 U/ml (Gibco, Waltham, MA, USA). The next day, the undigested tissue pieces were removed by centrifugation, and the supernatant was plated on gelatine (1 g/l) pre-treated dishes in αMEM (Euroclone) supplemented with 10% defined FBS (GE-Healthcare, Chicago, IL, USA), non-essential amino acids (Euroclone), L-alanine-L-glutamine (Euroclone), penicillin/streptomycin (Euroclone), antibiotic-antimycotic, and gentamycin.
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