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Development of Phenotype MicroArrays for Measuring Energy Producing Pathways in Mammalian Cells
Author(s) -
Wiater Lawrence A.,
Siri Mark,
Huang Richard H.,
Bochner Barry R.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a99-c
Subject(s) - formazan , catabolism , biochemistry , chemistry , cell culture , metabolism , cell , metabolic pathway , phenotype , biology , gene , genetics
Phenotype MicroArray™ (PM) technology was developed initially for microbial cells as a phenomic approach to scanning the pathways and physiology of the cell. This technology has now been extended to assay energy producing pathways in mammalian cells. The PMs measure cell‐mediated reduction of a tetrazolium to a colored formazan product that is dependent on exogenously added substrates present in microplate wells. Currently, four PM panels for mammalian cells have been developed to assay 367 substrates that include carbohydrates and polysaccharides, carboxylic acids, and a variety of amino acids in mono‐ and di‐peptide forms. Although all cells reduce the tetrazolium in the presence of glucose and mannose, unique patterns of substrate‐stimulated dye reduction have been obtained for all cells lines tested, including liver (C3A and HepG2), colon (Colo205), lung (A549), prostate (PC‐3), and blood (HL‐60 and CEM) cells. Indeed, although C3A cells were derived from HepG2 cells, their dye reduction patterns are similar but distinguishable, indicating some differences in cellular metabolism. Using A549 in further studies, we found that the serum component and the presence or absence of amino acids had a dramatic effect on a number of these catabolic pathways, especially in wells containing glycogen and pyruvate. PMs provide a new way to scan cellular metabolism as it relates to nutrition, obesity, diabetes and cancer. It can be employed to “fingerprint” the action of drugs and hormones on catabolic pathways. In addition, the technology can monitor changes in cultured cells that may occur through passage or infection. Supported by NIH Grant 9 R44 MH074145