Bme amino acid solution 50x

To promote the formation of tissues, bYS-MSCs (25 to 34 days of gestation) were cultured in α-MEM culture medium (minimum essential medium, Gibco) supplemented with 10% fetal bovine serum (Gibco), BME amino acid solution 50x (Sigma-Aldrich), MEM nonessential amino acids 100x (Sigma-Aldrich), β-mercaptoethanol solution 100x (Gibco), and a 1% v/v antibiotic solution (penicillin G 10.000 units/mL and streptomycin 10.000 μg/mL) and were trypsinized (TrypLE Express; Gibco-BRL). When they reached 80% confluence, they were replaced. From passage 4, the cells started an aggregation process to spontaneously form a three-dimensional tubular structure. To evaluate the tubular structure, histology, transmission electron microscopy, and electronic microscopy techniques were performed.

YS cells were cultured following the protocol of Mançanares et al. [1 (link)]. Cells were first washed in a solution of PBS-L (without calcium or magnesium) supplemented with antibiotics (5% penicillin-streptomycin; Invitrogen, Carlsbad, CA, USA) and then macerated enzymatically with 0.5% collagenase IV (Sigma-Aldrich) for 1 hour. The collagenase was inactivated with α-MEM culture medium (minimum essential medium, Gibco) supplemented with 10% fetal bovine serum (Gibco), BME amino acid solution 50x (Sigma-Aldrich), MEM nonessential amino acids 100x (Sigma-Aldrich), β-mercaptoethanol solution 100x (Gibco), and a 1% v/v antibiotic solution (penicillin G 10.000 units/mL and streptomycin 10.000μg/mL). Cells were plated in 25 cm² plates in a 37°C incubator with 5% CO₂.