Technical Abstract: Three Scutellaria species (Scutellaria lateriflora, S. costaricana and S. baicalensis) were grown in different in vitro physical environments: agar, liquid culture, and liquid culture with fiber-supported paper (with initial media volumes of 20 mL and 30 mL). During an eight-week time course, tissue growth was assessed and for each species, fresh weight (FW), dry weight (DW), % dry weight and multiplication ratio, along with water use and hyperhydricity, were compared. For S. lateriflora, plantlets grown in liquid were hyperhydric despite the greatest accumulation of dry mass, and multiplication diminished with time, as plants became hyperhydric. On the other hand, S. costaricana and S. baicalensis plantlets had higher FW and DW on agar. With all Scutellaria species tested, plantlets grown on agar or fiber-supported paper were not hyperhydric, and fiber-supported paper with 20 mL initial volume had plants with greatest %DW. The lowered uptake of water per plant was related to lowered hyperhydricity. The flavonoids baicalin, baicalein and wogonin were quantified in plants grown on fiber-supported paper culture. The baicalin concentrations in in vitro cultured S. lateriflora shoots was comparable to those of field-grown plants. The in vitro method presented a unique opportunity to enhance baicalein content and produce wogonin-rich roots. S. costaricana plantlets in vitro showed high levels of the three flavonoids, compared to S. baiclalensis and S. lateriflora. Growing non-hyperhydric tissues on fiber-supported paper, in vitro, allowed the clonal propagation of Scutellaria species with increased flavonoid content to proceed in a simple, controlled environment.