Development of an anti-metastatic assay and cytotoxicity test using a human cell line for screening natural products
2002
Rochanaruangrai, S.(National Science and Technology Development Agency, Bangkok (Thailand). National Center for Genetic Engineering and Biotechnology)
Two additional tests are being developed in the Bioassay laboratory at BIOTEC to increase it's capacity as a bioscreening service laboratory. Anit-metastatic assay, a screening test for compounds that specifically target invading tumours, is being developed based on the transwell invasion assay of Khutson et. Al. (1996). We found that the number of HT-1080 cells (fibrosarcoma) and the incubation time, which yields the highest number of invading cells is 2*10**(5) cells/well and longer than 6 hours, respectively. Currently, we are optimizing the assay conditions to allow quantification of cells by measuring MTT metabolite, instead of cell counting method used in the traditional assay, which is inaccurate and tedious. In vitro cytotoxicity tests for predicting human toxicity have been developed as alternatives of animal testing, which is laborious and very expensive. One cytotoxicity test that has been shown to give a good estimation of toxicity compared to results from animal tests is to measure ATP content at the endpoint as a marker for the viability of HL-60 cells. We plan to modify the HL-60 cytotoxicity test into a 96-well format from the original test tube method for higher throughput screening. At the moment, we found that 5*10**(4) HL-60 cells/well provide sufficient sensitivity for 96 well format testing.
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