Project Objective

The possibility of using this protein as a biopharmaceutical treatment for obesity is being investigated in this research. The oxygen uptake rate of the adipose cells was used as key measure of cell metabolism. The cells’ oxygen uptake rates are being measured to compare their fat metabolism. Preadipocytes and fully differentiated adipocytes of our wild-type (3T3-L1 mouse adipose cell line) and genetically altered cells that produce UCP1 (UCP1 cell line) were tested.

Hypothesis

The hypothesized results are that the cells transfected to produce UCP1 should have a higher oxygen uptake rate than the wild-type 3T3-L1 cells.

Figure 2 Oxygen Uptake Rate Measurement System

System

The cell culture flasks were filled with medium and a small stir bar and closed with a cap with the oxygen probe attached. The flask was placed on a stir plate enabling the medium to flow within the flask. The probe is attached to an amplifier which sends the signal through a National Instruments Data Acquisition Board relaying the signal to the computer. LabVIEW is used to control and collect the data which can then be used for further analysis.

Data Analysis

The raw data is collected through the LabVIEW program for one and a half to two hours. After data collection is complete, another LabVIEW program, written to analyze the data, crops the data to show the initial slope over the first ten minutes after the first minute was cropped off. The first 75 seconds are cropped to allow enough time for the oxygen probe to equilibrate to its new environment in the medium. A smoothing filter is then employed to reduce the noise in the data and fit to a linear curve.

Normalizing Data

It is important to normalize the data for each slope because the units measured are in percent per second, an arbitrary measurement. A DNA assay machine is used to count the number of total double-stranded pieces of DNA present in each cell culture sample after experimentation. Using a given number of DNA strands per cell the number of cells in each cell culture sample can be calculated. Using conversion factors the slopes can be normalized to mmHg/cells/second and finally parts per million (ppm) / cells / second.