Artificial urine (AU) is widely used for in vitro cellular study to simulate normal physiological environment of the kidney and urinary tract. However, some compositions of many of previously established AU formulas are out of their physiological ranges in normal human urine. Therefore, we established a new AU formula, named “AU-Siriraj,” and then performed systematic comparisons of AU-Siriraj with other six previously established AU formulas, ultrafiltrated (UF) urine, and blank–control to determine their compatibility with MDCK cells. The data indicate that AU-Siriraj is the best, and AU-3 is the second best, AU formula for in vitro cellular study.