Institute of Biology and Medicine, Wuhan University of Science and Technology, Huangjiahu Campus, Wuhan, Hubei Province, China.
School of Chemistry and Chemical Engineering, Wuhan University of Science and Technology, Qingshan Campus, Wuhan, Hubei Province, China.
PLoS One. 2017 Apr 18;12(4):e0176120. doi: 10.1371/journal.pone.0176120. eCollection 2017.
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.
哺乳动物细胞是非常重要的实验材料，广泛应用于生物和医学研究领域。为了满足各种研究需求，常常需要将哺乳动物细胞从一个实验室运输到另一个实验室。尽管传统的细胞运输方法可以维持高存活率，但却很繁琐且成本高昂。在本研究中，我们旨在通过研究不同温度处理的不同细胞系的存活率，开发一种简单且低成本的细胞运输方法。结果表明，与 16°C 或 22°C 相比，在 1°C 或 5°C 孵育的哺乳动物细胞的存活率显著降低。集落形成实验表明，与 16°C 或 22°C 相比，将哺乳动物细胞保存在 1°C 或 5°C 会导致恢复能力较差。增殖和凋亡实验的数据证实，M2 细胞在 16°C 或 22°C 下可以继续增殖，但在 1°C 或 5°C 下大量凋亡。低温处理后的哺乳动物细胞形态与未经处理的细胞几乎没有区别。定量 RT-PCR 和碱性磷酸酶染色证实，低温处理不会改变小鼠胚胎干细胞的特性。一项案例研究表明，直接悬浮在培养基中的哺乳动物细胞能够进行长距离运输，并保持高存活率和恢复能力。我们的研究结果不仅拓宽了对低温对哺乳动物细胞活力影响的认识，还为细胞运输提供了一种替代方法。
