Development of a PC12 cell cultivation and monitoring system for neuronal-like study

A variety of cell lines are often used to simulate the studies associated with the cell products, genetics, cellcell interaction, environmental interaction, intracellular flux and intracellular activity. Among these cell lines, the pheochromocytoma (PC12 for short) cell line is the one that is widely applied in in vitro study for investigating the neuronal apoptosis, neurochemistry, and neuronal differentiation. The development of an appropriate cell cultivation platform as well as the real-time image acquisition and analysis system becomes an important issue. The purpose of this paper is to develop a long-term and low-cost incubation system, including microscopic image analysis system for morphological analysis. The proposed incubation system can acquire cell image for analysis during cell cultivation. From the experimental results, the temperature of incubation system is found to be in the range of 37.3 ± 1.3°C, and the lower levels of the relative humidity vary within 89.7 ± 1.8%. The CO₂ concentration is controlled at 5.29 ± 0.71%. The cell number is increased from 1.00 × 10 ⁵ to 4.25 × 10⁵ cells/ml after 48 h of culture. The cell viability test shows that this developed incubation system works well for PC12 cells cultivation; it can also acquire and segment the cell image during the cell cultivation. Besides, in the image analysis software design, the region-growing and modified blob labeling techniques are also employed to segment the PC12 cell image under varied culture conditions for later morphological and feature quantification analysis.