AI Article Synopsis

  • Multiple myeloma (MM) is a type of cancer affecting plasma cells in the bone marrow, characterized by altered metabolism, particularly in glycolysis, lipid metabolism, and mitochondrial respiration.
  • Recent advancements in technology, like the Seahorse™ XF analyzer, allow researchers to monitor real-time changes in cell metabolism, specifically glycolytic flux and mitochondrial activity, in various cell types, though non-adherent suspension cells present challenges.
  • The authors present an optimized protocol for analyzing live myeloma cells under drug treatment using the XF analyzer, demonstrating that their method yields consistent results, which could be beneficial for future research on metabolic processes in similar cells.

Article Abstract

Multiple myeloma (MM) is an incurable plasma cell malignancy primarily localized within the bone marrow (BM). Myeloma plasma cells, like many other cancer cells, change their metabolism in response to internal and external stimuli. The main metabolic alterations of MM cells include deregulated glycolysis (commonly associated with enhanced uptake and utilization of glucose), lipid metabolism dysregulation, as well as deregulated mitochondrial respiration (commonly associated with the deregulated formation of reactive oxygen species). Over the past decade, the discovery of novel methodologies and the commercialization of sophisticated instrumentation and reagents have facilitated the detection of real-time changes in cellular bioenergetics. Of those, the Seahorse™ extracellular flux (XF) analyzer has been widely used to evaluate the glycolytic flux and mitochondrial respiration in many cell types. While adherent cell lines are easy to use with this technology, non-adherent suspension cells are more difficult to handle especially when their metabolic activities are being investigated in response to drug treatment. Here, we provide an integrated protocol that allows the detection of extracellular acidification rate (ECAR) of live myeloma plasma cells in response to chemotherapeutic drugs. Our optimized protocol consists of treating myeloma cells with cytotoxic drug of interest in a standard culture plate prior to the real-time analysis in the XF analyzer. Furthermore, we provide results of experiments in which the metabolic activities of myeloma cells in response to cytotoxic treatment were compared between the manufacturer's basic procedure and our optimized protocol. Our observations suggest that our integrated protocol can be used to achieve consistent, well-standardized results and thus it may have broad applications in studies focusing on the characterization of metabolic events in non-adherent suspension cells.

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Source
http://dx.doi.org/10.1007/978-1-0716-3247-5_21DOI Listing

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