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Biosensors have emerged as promising tools for the analysis of single-cell growth. This study focuses on the fabrication of an impedance biosensor to monitor the growth of NIH/3T3 cells by measuring changes in their capacitance over time. The results showed an increase in capacitance corresponding to the growth of the cells. However, it was observed that the capacitance behaved differently in the early (0–2.33 h) and late (2.33–48 h) stages of cell growth. In the early stage, the cells in the medium were still in a sinking state, whereas in the late stage, most of the cells were in contact with the sensor surface, undergoing growth and division. As a result, the late-stage capacitance was higher than the early stage capacitance. The average capacitance values of a single cell were recorded at different frequencies and showed the same phenomenon at all frequencies.