Induced pluripotent stem cells can utilize lactate as a metabolic substrate to support proliferation
Induced pluripotent stem cells can utilize lactate as a metabolic substrate to support proliferation
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DOI：
10.1002/btpr.3090
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发表时间：
2020-11-04
影响因子：
2.9
通讯作者：
Harcum, Sarah W.
中科院分区：
文献类型：
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作者：
Odenwelder, Daniel C.;Lu, Xiaoming;Harcum, Sarah W.
人类诱导多能干细胞（iPSC）有望改善基于细胞的疗法。然而，为了满足不断增长的需求并成为临床上有影响力的，必须大量生产足够的高质量iPSC，这是一项超出当前能力的任务。在这项研究中，使用平行标记代谢通量分析（C-13-MFA）检查K3 iPSCs培养物，以量化相关生物加工阶段的细胞内通量：在推注葡萄糖进料之前和之后，代表初始培养基浓度的葡萄糖浓度和代表补料分批培养条件的高乳酸盐浓度。葡萄糖和乳酸浓度也代表了在3D细胞聚集体内不同位置可能遇到的浓度。此外，开发了一种新的方法，允许同位素示踪剂[U-C-13（3）]乳酸盐用于C-13-MFA模型。结果表明，高胞外乳酸浓度降低葡萄糖消耗和乳酸产生，而葡萄糖浓度本身并不影响有氧糖酵解速率。此外，对于高乳酸盐培养物，乳酸盐用作代谢底物以支持氧化线粒体代谢。这些结果表明，iPSC具有代谢灵活性，并且具有代谢乳酸盐以支持指数生长的能力，并且单独的高乳酸盐浓度不会对iPSC增殖产生不利影响。
Human-induced pluripotent stem cells (iPSCs) hold the promise to improve cell-based therapies. Yet, to meet rising demands and become clinically impactful, sufficient high-quality iPSCs in quantity must be generated, a task that exceeds current capabilities. In this study, K3 iPSCs cultures were examined using parallel-labeling metabolic flux analysis (C-13-MFA) to quantify intracellular fluxes at relevant bioprocessing stages: glucose concentrations representative of initial media concentrations and high lactate concentrations representative of fed-batch culture conditions, prior to and after bolus glucose feeds. The glucose and lactate concentrations are also representative of concentrations that might be encountered at different locations within 3D cell aggregates. Furthermore, a novel method was developed to allow the isotopic tracer [U-C-13(3)] lactate to be used in the C-13-MFA model. The results indicated that high extracellular lactate concentrations decreased glucose consumption and lactate production, while glucose concentrations alone did not affect rates of aerobic glycolysis. Moreover, for the high lactate cultures, lactate was used as a metabolic substrate to support oxidative mitochondrial metabolism. These results demonstrate that iPSCs have metabolic flexibility and possess the capacity to metabolize lactate to support exponential growth, and that high lactate concentrations alone do not adversely impact iPSC proliferation.