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首页> 外文期刊>Artificial Organs >Labeling of Primary Human Hepatocytes With Micron-Sized Iron Oxide Particles in Suspension Culture Suitable for Large-Scale Preparation
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Labeling of Primary Human Hepatocytes With Micron-Sized Iron Oxide Particles in Suspension Culture Suitable for Large-Scale Preparation

机译：适用于大规模制备的悬浮培养中的微米级氧化铁颗粒标记人类原代肝细胞

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Labeling of hepatocytes with micron-sized iron oxide particles (MPIOs) enables cell detection using clinical magnetic resonance equipment. For clinical applications, large numbers of cells must be labeled in a simple and rapid manner and have to be applied in suspension. However, all existing protocols are based on adhesion culture labeling with subsequent resuspension, only suitable for small experimental settings. The aim of this study was to investigate the feasibility of preparing MPIO-labeled primary human hepatocytes in a temporary suspension culture. Human hepatocytes were isolated from 16 donors and labeled with MPIOs in suspension, using the Rotary Cell Culture System. Particle incorporation was investigated by light and electron microscopy. Cells were compared with adhesion culture-labeled and subsequently enzymatically resuspended cells. During a period of 5 days, hepatocyte-specific parameters of cell damage (aspartate aminotransferase and alanine aminotransferase) and metabolic activity (urea and albumin) were analyzed (n?=?7). Suspension cultures showed a higher outcome in cell recovery compared with the conventional labeling method. When incubated with 180 particles/viable cell for 4?h, the mean particle uptake was 28.8 particles/cell at a labeling efficiency of 95.1%. Labeling in suspension had no adverse effects on cell integrity or metabolic activity. We conclude that labeling of human hepatocytes in suspension is feasible and simple and may serve future large-scale processing of cells.
机译：用微米级氧化铁颗粒（MPIO）标记肝细胞，可以使用临床磁共振设备进行细胞检测。对于临床应用，必须以简单，快速的方式标记大量细胞，并且必须将其以悬浮液形式使用。但是，所有现有方案均基于粘附培养标记并随后重悬，仅适用于小型实验环境。这项研究的目的是研究在临时悬浮培养物中制备MPIO标记的原代人肝细胞的可行性。使用旋转细胞培养系统从16个供体中分离出人类肝细胞，并用悬浮的MPIOs进行标记。通过光学和电子显微镜研究了颗粒结合。将细胞与粘附培养标记的并随后酶促重悬的细胞进行比较。在5天的时间内，分析了细胞损伤的肝细胞特异性参数（天冬氨酸转氨酶和丙氨酸转氨酶）和代谢活性（尿素和白蛋白）（n == 7）。与常规标记方法相比，悬浮培养显示出更高的细胞回收率。当与180个颗粒/活细胞孵育4？h时，平均颗粒吸收率为28.8个颗粒/细胞，标记效率为95.1％。悬浮液中的标记对细胞完整性或代谢活性没有不利影响。我们得出的结论是，在悬浮液中标记人肝细胞是可行且简单的，可能会为将来的细胞大规模加工提供服务。

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