The analysis of viability for mammalian cells treated at different temperatures and its application in cell shipment

Abstract

Mammalian cells are very important experimental materials and widely used in biological and medical research fields. It is often required that mammalian cells are transported from one laboratory to another to meet with various researches. Conventional methods for cell shipment are laborious and costive despite of maintaining high viability. In this study we aimed to develop a simple and low-cost method for cell shipment by investigating the viabilities of different cell lines treated at different temperatures. We show that the viability of mammalian cells incubated at 1°C or 5°C significantly reduced when compared with that at 16°C or 22°C. Colony formation assays revealed that preservation of mammalian cells at 1°C or 5°C led to a poorer recovery than that at 16°C or 22°C. The data from proliferation and apoptotic assays confirmed that M2 cells could continue to proliferate at 16°C or 22°C, but massive death was caused by apoptosis at 1°C or 5°C. The morphology of mammalian cells treated under hypothermia showed little difference from that of the untreated cells. Quantitative RT-PCR and alkaline phosphatase staining confirmed that hypothermic treatment did not change the identity of mouse embryonic stem cells. A case study showed that mammalian cells directly suspended in culture medium were able to be shipped for long distance and maintained a high level of viability and recovery. Our findings not only broaden the understanding to the effect of hypothermia on the viability of mammalian cells, but also provide an alternative approach for cell shipment.

Effects of Temperatures and Basal Media on Primary Culture of the Blastomeres Derived from the Embryos at Blastula Stage in Marine Medaka Oryzias dancena

“…In cell culture, the optimal temperature depends on the body temperature of the animals from which the cells were derived or the anatomic difference (Freshney, 2010). Therefore, the cells and tissues from human and mouse are usually cultured in 37°C according to their body temperature (Wang et al, 2017). But, in some cases like testis tissues, they are cultured in less than 37°C based on the anatomic difference (Gohbara et al, 2010).…”

Transfection types, methods and strategies: a technical review

“…Another interesting observation was that 37 C was the optimal incubation temperature that could help reach higher transfection efficiency in primary cells (Young et al, 2002). This phenomenon could be because 37 C is the optimal culture temperature for mammalian cells (Wang et al, 2017a). Meanwhile, chemical transfection appeared to be less appealing than viral and physical transfection for transfecting primary cells, especially in human primary stem cells.…”

Human Islet Isolation and Distribution Efforts for Clinical and Basic Research

“…Exposure of islets to temperatures beyond their physiological range for an extended period of time negatively impacts their quality. It has been shown that hypothermic conditions (1-4 °C) drastically reduce the viability of mammalian cells [54], while high temperatures (43 °C) decrease the survival rates of early xenografts of pig islets in mice [55]. The Integrated Islet Distribution Program (IIDP) provides ColdMark ® and WarmMark ® indicators to detect undesirable temperature changes.…”