Scutellaria baicalensis Georgi (SJ) is a perennial plant and its root has been used in oriental traditional medicine for treatment of fever, inflammation, diarrhea and anticancer effect, etc. In this study, plant tissue culture system for SJ was developed. Stem piece of younger plant was optimum explant for callus induction and growth on MS medium supplemented with NAA 1.0 ㎎/L plus BA 0.5 ㎎/L. Plantlet regeneration through callus culture was well on MS medium containing NAA 1.0 mg/L. SJ has been known biologically active substances such as baicalin, baiclein, and wogonin. This study was carried out to examine the effect of plant growth regulators for production of baicalin, baicalein, and wogonin through callus culture. The HPLC pattern of callus extract was identical to that of standard solution, it shows that the callus produced by tissue culture has the same flavonoids composition of SJ. Baicalin, baicalein, and wogonin production was 471.5~52.8 ㎍/g, 137.6~4.0 ㎍/g, and 16.6~1.3 ㎍/g, respectively, on MS media with nine different plant growth regulator combinations. This may indicate that plant tissue culture of SJ possible to produce the biologically active substances effectively.