Abstract: This disclosure relates to fluorescent cell lines and to the use of such cell lines in monitoring cellular activity, such as angiogenesis. This disclosure further relates to the use of such cell lines in a three-dimensional cell culture to monitor angiogenic and metastatic potential of tumor cells and selecting personalized therapeutics for treatment of cancer.

Abstract: Disclosed is an in vitro assay for evaluating cellular activity, in which a culture is provided that contains one or more histologically different isolated mammalian cell-line that stably and constitutively express fluorescent proteins having a different emission spectra. The culture is assessed for cellular activity by quantifying fluorescence or detecting a pattern of fluorescence from fluorescent proteins present in the culture. In some embodiments, the cellular activity of the cell-lines is assessed by determining one or more of the growth rate, migration potential, or tubule formation potential of the cell-lines using the quantified fluorescence or pattern of fluorescence. Also disclosed are cell-lines have been stably transfected with mammalian expression plasmids that constitutively express different fluorescent proteins. Kits for performing the disclosed assays, which include the disclosed fluorescent cell-lines, are also disclosed.

Abstract: The present invention provides a new protocol for quantifying multiplex real-time polymerase chain reaction (PCR). In particular, the present invention provides methods of quantifying multiple PCR products or amplicons in a single real-time PCR reaction based on the different melting temperatures (Tm) of each amplicon and the emission changes of double stranded DNA dyes such as SYBR Green I when amplicons are in duplex or in separation. For a specific amplicon with a Tm, the emission difference between the emission reading taken at a temperature below the Tm and the emission reading taken at a temperature above the Tm corresponds to the emission value of the amplicon in duplex. Accordingly, the emission difference of each amplicon in a single PCR reaction can be used to quantify each amplicon. The present invention further provides computer programs or computer products which perform the methods described herein.