Oca-Cossio J, Simpson N E, Han Z, Stacpoole P W, Constantinidis I
Department of Medicine, Division of Endocrinology, University of Florida, Gainesville, FL 32610-0226, USA.
J Mater Sci Mater Med. 2005 Jun;16(6):521-4. doi: 10.1007/s10856-005-0527-y.
The long-term objective of our research is to study the biochemical consequences of primary genetic defects of the Pyruvate Dehydrogenase Complex, a key mitochondrial enzyme complex, by NMR spectroscopy. An established method to obtain energetic and metabolic information from intact cells involves the use of 31P and 13C NMR spectroscopic techniques. NMR spectra from live and fully functional cells can be obtained from cells encapsulated within alginate beads and maintained in a perfusion bioreactor throughout the NMR experiment. However, before spectroscopic studies can commence, the effects of alginate encapsulation on the general metabolism and mitochondrial activity of fibroblasts need to be determined. in this study we report glucose consumption and flow cytometry measurements (with the fluorescent markers MitoTracker GreenFM and Nonyl-acridine Orange to determine the mitochondrial status and mass) of healthy human fibroblasts encapsulated in a mannuronic acid-rich alginate matrix. The results show that alginate encapsulation of fibroblasts does not affect the glucose consumption, the mitochondrial integrity, or the mitochondrial mass during 21 days of in vitro culture.
我们研究的长期目标是通过核磁共振光谱法研究丙酮酸脱氢酶复合体（一种关键的线粒体酶复合体）原发性基因缺陷的生化后果。一种从完整细胞中获取能量和代谢信息的既定方法涉及使用³¹P和¹³C核磁共振光谱技术。在整个核磁共振实验过程中，可以从包裹在藻酸盐珠粒中并维持在灌注生物反应器中的细胞获得来自活的且功能完全正常的细胞的核磁共振光谱。然而，在开始光谱学研究之前，需要确定藻酸盐包封对成纤维细胞一般代谢和线粒体活性的影响。在本研究中，我们报告了包裹在富含甘露糖醛酸的藻酸盐基质中的健康人成纤维细胞的葡萄糖消耗和流式细胞术测量结果（使用荧光标记物MitoTracker GreenFM和壬基吖啶橙来确定线粒体状态和质量）。结果表明，在体外培养21天期间，藻酸盐对成纤维细胞的包封不会影响葡萄糖消耗、线粒体完整性或线粒体质量。
