Methods and kits for obtaining a metabolic profile of living animal cells

A method for observing effects of a candidate compound on substrate utilization by animal cells, comprising the steps of:

• a) providing:
• i) a suspension medium comprising at least one animal cell;
• ii) one or more testing devices comprising a positive control well and a negative control well;
• iii) said one or more testing devices further comprising a plurality of testing wells and wherein each of said plurality of testing wells comprises a plurality of the same testing substrates wherein each of said plurality of testing wells and said positive control well contain at least one testing substrate;
• and
• iv) a redox indicator;
• b) introducing a first portion of said suspension medium into said positive control well and said negative control well and incubating said positive control well and said negative control well;
• c) introducing a first portion of said redox indicator into said positive control well and said negative control well wherein an adaptation period length is determined by a greater measurable color developed in said positive control well as compared to measureable color developed in said negative control well;
• d) introducing a second portion of said suspension medium into said plurality of testing wells;
• e) introducing a candidate compound at a range of concentrations into said plurality of testing wells;
• f) pre-incubating said second portion of said suspension medium in said plurality of testing wells for said adaptation period length;
• g) introducing a second portion of said redox indicator into said plurality of testing wells;
• h) incubating said second portion of said suspension medium with said second portion of said redox indicator wherein a color development occurs in at least one of said plurality of testing wells;
• i) measuring said color development to obtain a metabolic profile of said second portion of said suspension medium, wherein said color development is selected from the group consisting of increased color development and decreased color development;
• and
• j) analyzing said metabolic profile to determine phenotypes of said animal cells wherein an utilization effect of said candidate compound is identified based on said testing substrates.

A method for phenotyping a multiplexed metabolic profile of animal cells, comprising the steps of:

• a) providing a testing device comprising a plurality of testing wells and a plurality of testing substrates, wherein each of said testing wells contains at least one unique testing substrate;
• b) suspending multiple samples comprising animal cells in multiple suspension media to produce multiple suspensions, wherein said multiple suspension media are selected from the group consisting of three or more of a low serum medium, an intermediate serum medium, a high serum medium, a whole serum medium, a dialyzed serum medium, an amino acid-rich medium, and an amino acid poor-medium, wherein said multiple suspension media are aqueous solutions comprising extraneous substrates;
• c) introducing said multiple suspensions of animal cells into said testing wells of said testing device;
• d) pre-incubating said animal cells in said testing device wherein said extraneous substrates are depleted;
• e) introducing a redox indicator into said plurality of testing wells that have been depleted of said extraneous substrates in step d) of said testing device;
• f) incubating said animal cells in said testing device wherein a color development occurs in at least one of said each of said plurality of testing wells primarily due to utilization of said at least one unique testing substrate;
• g) measuring said color development to obtain a multiplexed metabolic profile of said animal cells, comprising multiple metabolic profiles;
• and
• h) analyzing said multiple metabolic profiles to determine phenotypes of said animal cells.