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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
The differentiation capacity of the cells has been tested using the differentiation protocol described in Xue et al (Nature Medicine 2015). Oil Red O staining showed that the majority of cells were differentiated into lipid-laden adipocytes. Gene expression analysis demonstrated differentiated A41 hBAT expressed high level of adipocyte markers such as PPARg, FABP4, and brown fat-specific marker UCP1.
To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.
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Xue R, et al. Clonal analyses and gene profiling identify genetic biomarkers of the thermogenic potential of human brown and white preadipocytes. Nat Med 21(7):760-768, 2015. PubMed: 26076036
Shamsi F, Tseng YH. Protocols for Generation of Immortalized Human Brown and White Preadipocyte Cell Lines. Methods Mol Biol 1566:77-85, 2017. PubMed: 28244042