Hwang Emma Y, Pappas Dimitri, Jeevarajan Antony S, Anderson Melody M
Biotechnology Program, Wyle Laboratories, Houston, TX, USA.
Biomed Microdevices. 2004 Sep;6(3):241-9. doi: 10.1023/B:BMMD.0000042054.02940.b6.
Compact and automated sensors are desired for assessing the health of cell cultures in biotechnology experiments. While several single-analyte sensors exist to measure culture health, a multi-analyte sensor would simplify the cell culture system. One such multi-analyte sensor, the Paratrend 7 manufactured by Diametrics Medical, consists of three optical fibers for measuring pH, dissolved carbon dioxide (pCO(2)), dissolved oxygen (pO(2)), and a thermocouple to measure temperature. The sensor bundle was designed for intra-vascular measurements in clinical settings, and can be used in bioreactors operated both on the ground and in NASA's Space Shuttle and International Space Station (ISS) experiments.
A Paratrend 7 sensor was placed at the outlet of a bioreactor inoculated with BHK-21 (baby hamster kidney) cells. The pH, pCO(2), pO(2), and temperature data were transferred continuously to an external computer. Cell culture medium, manually extracted from the bioreactor through a sampling port, was also assayed using a bench top blood gas analyzer (BGA).
Two Paratrend 7 sensors were used over a single cell culture experiment (64 days). When compared to the manually obtained BGA samples, the sensor had good agreement for pH, pCO(2), and pO(2) with bias (and precision) 0.005(0.024), 8.0 mmHg (4.4 mmHg), and 11 mmHg (17 mmHg), respectively for the first two sensors. A third Paratrend sensor (operated for 141 days) had similar agreement (0.02+/-0.15 for pH, -4+/-8 mm Hg for pCO(2), and 24+/-18 mmHg for pO(2)).
The resulting biases and precisions are com- parable to Paratrend sensor clinical results. Although the pO(2) differences may be acceptable for clinically relevant measurement ranges, the O(2) sensor in this bundle may not be reliable enough for the ranges of pO(2) in these cell culture studies without periodic calibration.
在生物技术实验中,需要紧凑且自动化的传感器来评估细胞培养的健康状况。虽然存在几种用于测量培养健康状况的单分析物传感器,但多分析物传感器将简化细胞培养系统。一种这样的多分析物传感器,即由 Diametrics Medical 制造的 Paratrend 7,由三根用于测量 pH、溶解二氧化碳(pCO₂)、溶解氧(pO₂)的光纤和一个用于测量温度的热电偶组成。该传感器束设计用于临床环境中的血管内测量,并且可用于在地面以及美国国家航空航天局(NASA)的航天飞机和国际空间站(ISS)实验中运行的生物反应器。
将一个 Paratrend 7 传感器放置在接种了 BHK - 21(幼仓鼠肾)细胞的生物反应器出口处。pH、pCO₂、pO₂ 和温度数据被连续传输到外部计算机。还使用台式血气分析仪(BGA)对通过采样端口从生物反应器中手动提取的细胞培养基进行了分析。
在一次细胞培养实验(64 天)中使用了两个 Paratrend 7 传感器。与手动获取的 BGA 样本相比,对于前两个传感器,该传感器在 pH、pCO₂ 和 pO₂ 方面具有良好的一致性,偏差(和精密度)分别为 0.005(0.024)、8.0 mmHg(4.4 mmHg)和 11 mmHg(17 mmHg)。第三个 Paratrend 传感器(运行 141 天)具有类似的一致性(pH 为 0.02±0.15,pCO₂ 为 -4±8 mmHg,pO₂ 为 24±18 mmHg)。
所得偏差和精密度与 Paratrend 传感器的临床结果相当。尽管对于临床相关测量范围,pO₂ 差异可能是可接受的,但在这些细胞培养研究中,该束中的 O₂ 传感器在没有定期校准的情况下,对于 pO₂ 范围可能不够可靠。