The Effect of Time and Holding Media on Cell Derivation of Balb/C Mouse at Room Temperature

Abstract

Efficient cell derivation from post-mortem tissues is critical for biomedical research, yet its success is often compromised by delayed processing and inadequate preservation. This study aimed to assess the impact of post-mortem time intervals and the use of holding media on the success rate of cell derivation from Balb/C mice tissues at room temperature. Tissue samples were divided into four groups based on time (0 hours, 6 hours, 12 hours, and 24 hours post-mortem) and whether they were retained in media or not. Then, the samples were processed by washing, mincing, neutralising, centrifuging at 1000 rpm for 5 minutes, and resuspending before plating in 12-well plates in triplicates. The data were analysed using RStudio. Tissues retained in media maintained viable cell morphology and a gradual decline in plating efficiency up to 24 hours (75%), whereas tissues without media showed loss of viability as early as 6 hours, with plating efficiency dropping to 8.33% and declining further over time. Statistical analysis revealed a p-value of 0.283 for tissues retained in media, indicating no significant difference in viability over time, whereas tissues without media had a p-value <0.001, indicating a significant time-dependent decline. The time factor alone showed no significant impact on derivation success for tissues in media, maintaining tissue hydration prior to derivation is crucial for ensuring higher viability and successful outcomes. It is hoped that this study could serve as a foundation for future research, further advancing efforts and activities related to planetary health.