Thymidine bioassay

The in vitro mitogenic bioassay used to determine the biological activity of KGF was similar to that described by Rubin, et al (13). The assay measured the incorporation of [ H]-thymidine by Balb/MK epidermal keratinocytes. 

IL-lra and TGF-pi on IL-1 induced activity in the coculture bioassay and on IL-2 induced CTLL proliferation, serial dilutions of IL-ip (0.78-50pg/mL), IL-2 (0.025-1.6U/mL), IL-lra (1-100ng/mL), TGF-pi (0.01-10ng/mL), and test compound in TCM supplemented with 10% FCS are used. Overall, 50-pL aliquots of each dilution of IL-lra and IL-ip are dispensed and pipetted into 96-well microtiter plates, respectively, and 50pL of the suspension of ELA4.NOB-1 and CTLL cells are ali-quoted into each well, incubated, and treated with H-thymidine as described above similar assays are set up for IL-lra and IL-2, for TGF-pi and IL-ip, and for TGF-pl and IL-2. 

Figure 5.14 Effects of increasing concentrations of falcarinol on proliferation, measured by incorporation of [methyl- H]thymidine into mammary epithelial cells prepared from prepubertal Friesian heifers and grown in 3-dimensional collagen gels (redrawn from Hansen efa/.2003). Stimulatory as well as inhibitory effects on cell proliferation were significantly different from those obtained in basal medium (BM). No effect on proliferation was observed for p-carotene when tested in the same bioassay (Hansen etal. 2003).

---