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.
Highlights
Mammalian cells including primary cells and cell lines are very important experimental materials and extensively utilized in the research field of biological and medical sciences
A pilot experiment was initially performed with human melanoma cells (M2). 3Ã105 of M2 cells were suspended with 1 mL of culture medium within a 2-mL Eppendorf microtube, followed by hypothermic treatment and stained by Fluoresein diacetate (FDA) and propidium iodide (PI) solution; the stained cells were subjected to statistical analysis (Fig 1A)
All of the cell lines tested above were stored in their own culture medium; we examined whether mammalian cells can survive longer when stored in commercial preservation medium under hypothermia
Summary
Mammalian cells including primary cells and cell lines are very important experimental materials and extensively utilized in the research field of biological and medical sciences. It is inevitable that the mammalian cells have to be shipped from one laboratory to another to meet with various researches around the world. Conventional method for cell shipment is that cryopreserved cells are transported with dry ice with in foam container; which shows little influence. An alternative method widely used by local companies or laboratories is directly to ship the cultured cells in the flask fully filled with cell culture medium [3, 4]; but the disadvantage of this method is not suitable for long-distance shipment [5]. Previous and recent studies showed that mammalian cells can be transported for long distance at ambient temperature by mixing the cells with agarose gel-or matrigel-based media [2, 6] and maintain a high rate of cell recovery after transportation for a few days. Whether mammalian cells can be shipped in a simple mode at ambient temperature remains unclear
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