Source: Shen, L., et al. Evaluation of Amino Acid Consumption in Cultured Bone Cells and Isolated Bone Shafts. J. Vis. Exp. (2022).
This video demonstrates an in vitro technique for estimating amino acid uptake by bone marrow-derived stromal cells. Na+-dependent amino acid transporters mediate the uptake of radiolabeled amino acids inside the cells, and the cellular uptake is estimated by measuring the radioactivity.
The use of radioactive materials (RAM) requires prior approval by a designated safety committee at each institution.
1. Amino acid uptake in cells
The authors have nothing to disclose.
0.25% trypsin | Gibco | 25200 | |
12-well plate | Corning | 3513 | |
Beckman LS6500 scintillation counter | |||
Calcium chloride | Sigma | C1016 | |
choline chloride | Sigma | C7077 | |
D-(+)-Glucose solution | Sigma | G8769 | |
DPBS | Gibco | 14190 | |
HEPES(1M) | Gibco | 15630 | |
L-[3,4-3H(N)]-Glutamine | PerkinElmer | NET551250UC | |
Liquid scintilation vials | Sigma | Z190535 | |
lithium chloride solution, 8M | Sigma | L7026 | |
Magnesium chloride | Sigma | M8266 | |
MEMα | Gibco | 12561 | |
Microcentrifuge tube, 15mL | Biotix | 89511-256 | |
Potassium chloride | Sigma | P3911 | |
Sodium bicarbonate | Sigma | S6014 | |
sodium chloride | Sigma | S9888 | |
Sodium dodecyl sulfate | Sigma | 436143 | |
Ultima Gold (Scintillation solution) | PerkinElmer | 6013329 | |
α-(Methylamino)isobutyric acid | Sigma | M2383 |