Chrest Brett R, Montgomery McLane M, Aruleba Raphael T, Krassovskaia Polina, Pacheco Emely A, Hagen James T, Vandiver Kayla J, Tung Kang, Alexander Molly K, Williamson Nicholas C, Taylor Joshua G, Bessetti Riley N, Belcher Heather A, Jevtovic Filip, Terwilliger Zoe S, Minchew Everett C, Zeczycki Tonya N, May Linda, Broskey Nicholas T, Geyer Christopher B, Litwa Karen, Spangenburg Espen E, Hannan Johanna L, Ellis Jessica M, McClung Joseph M, Neufer P Darrell, Fisher-Wellman Kelsey H
Department of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC, USA.
Department of Physiology, East Carolina University, Greenville, NC, USA.
Cancer Metab. 2025 May 26;13(1):25. doi: 10.1186/s40170-025-00395-1.
Increasing emphasis has been placed on improving the physiological relevance of cell culture media with formulations such as Human Plasma-Like Medium (HPLM). Given that shifts in mitochondrial metabolism and nutrient use are emerging as anti-cancer targets, the present study sought to investigate the impact of culture media formulation on mitochondrial bioenergetics and cancer cell growth. To do this, we used acute myeloid leukemia (AML) cells and compared acute and chronic effects of HPLM versus different supraphysiological medias. The AML mitochondrial phenotype was largely unaffected by exposure to either physiological or supraphysiological medias, establishing that the key features of AML mitochondria remain phenotypically stable under diverse nutrient conditions and proliferation rates. Both acute and chronic culturing in HPLM slowed AML cell proliferation. However, merely identifying and supplementing single nutrients that were deficient in HPLM did not improve proliferation and was not sufficient to pinpoint actionable fuel preferences. Transferring cells back to native Iscove's Modified Dulbecco's Medium (IMDM) media immediately restored the proliferative phenotype, suggesting responsiveness to the entirety of the nutrient environment. Supraphysiological culture medias other than IMDM were all characterized by slower proliferation; however, none were associated with changes in cell viability, demonstrating that the native culture medium is optimal if the experimental aim is maximal growth. Despite Dulbecco's Modified Eagle Medium (DMEM) being similar in nutrient composition to IMDM and categorized as supraphysiological, both acute and chronic culturing in DMEM resulted in slower growth, akin to what was observed with HPLM. Altogether, independent of growth, AML mitochondria remain largely unperturbed by changes in the culture media, and rather than specific nutrients or physiological relevance, AML cell proliferation is influenced by the complete nutrient profile.
人们越来越重视通过诸如类人血浆培养基（HPLM）等配方来提高细胞培养基的生理相关性。鉴于线粒体代谢和营养物质利用的变化正成为抗癌靶点，本研究旨在探讨培养基配方对线粒体生物能量学和癌细胞生长的影响。为此，我们使用急性髓系白血病（AML）细胞，比较了HPLM与不同超生理培养基的急性和慢性影响。AML线粒体表型在很大程度上不受生理或超生理培养基暴露的影响，这表明在不同的营养条件和增殖速率下，AML线粒体的关键特征在表型上保持稳定。在HPLM中进行急性和慢性培养均减缓了AML细胞的增殖。然而，仅仅识别和补充HPLM中缺乏的单一营养素并不能改善增殖，也不足以确定可操作的燃料偏好。将细胞立即转移回天然的伊斯科夫改良杜尔贝科培养基（IMDM）中可立即恢复增殖表型，这表明细胞对整个营养环境有反应。除IMDM外的超生理培养基均具有增殖较慢的特征；然而，没有一种与细胞活力的变化相关，这表明如果实验目的是实现最大生长，天然培养基是最佳选择。尽管杜尔贝科改良伊格尔培养基（DMEM）在营养成分上与IMDM相似且被归类为超生理培养基，但在DMEM中进行急性和慢性培养均导致生长较慢，这与在HPLM中观察到的情况类似。总之，与生长无关，AML线粒体在很大程度上不受培养基变化的干扰，并且AML细胞增殖受完整营养谱的影响，而不是特定营养素或生理相关性的影响。
