BACKGROUND: The objective of this work was to decellularize artificial tissue without using surfactant solutions. For this purpose, supercritical carbon dioxide was used as the extraction medium. RESULTS: Supercritical carbon dioxide containing a small amount of entrainer was a suitable medium to extract both cell nuclei and cell membranes from artificial tissue. Under gentle extraction conditions (15MPa, 37 degrees C), cell nuclei were satisfactorily extracted from tissue within 1 h. In contrast, the efficiency of phospholipid removal depended strongly on the transfer rate of carbon dioxide in the interior of the tissue. Mechanical strength of tissue was not decreased even with prolonged treatment. CONCLUSION: Acellular artificial tissues could be prepared quickly by treatment with a carbon dioxide/entrainer system. The prepared acellular tissue could be obtained in absolutely dry condition. This is advantageous from the viewpoint of long-term preservation without putrefaction and contamination. (c) 2008 Society of Chemical Industry.