Santoro R, Krause C, Martin I, Wendt D
Departments of Surgery and of Biomedicine, University Hospital Basel, Switzerland.
PreSens Precision Sensing GmbH, Regensburg, Germany.
J Tissue Eng Regen Med. 2012 Oct;6(9):696-701. doi: 10.1002/term.473. Epub 2011 Sep 20.
Regulatory guidelines have established the importance of introducing quantitative quality controls during the production and/or at the time of release of cellular grafts for clinical applications. In this study we aimed to determine whether on-line measurements of oxygen can be used as a non-destructive method to estimate the number of chondrocytes within an engineered cartilage graft. Human chondrocytes were seeded and cultured in a perfusion bioreactor, and oxygen levels in the culture medium were continuously monitored at the inlet and outlet of the bioreactor chamber throughout the culture period. We found that the drop in oxygen across the perfused construct was linearly correlated with the number of cells per construct (R(2) = 0.82, p < 0.0001). The method was valid for a wide range of cell numbers, including cell densities currently used in the manufacture of cartilage grafts for clinical applications. Given that few or no non-destructive assays that quantitatively characterize an engineered construct currently exist, this non-invasive method could represent a relevant instrument in regulatory compliant manufacturing of engineered grafts meeting specific quality criteria.
监管指南已明确了在临床应用的细胞移植物生产过程中和/或放行时引入定量质量控制的重要性。在本研究中，我们旨在确定在线氧气测量是否可作为一种非破坏性方法来估计工程化软骨移植物中的软骨细胞数量。将人软骨细胞接种并培养在灌注生物反应器中，在整个培养期间，连续监测生物反应器腔室入口和出口处培养基中的氧气水平。我们发现，灌注构建体中氧气的下降与每个构建体中的细胞数量呈线性相关（R² = 0.82，p < 0.0001）。该方法对广泛的细胞数量均有效，包括目前临床应用的软骨移植物制造中所使用的细胞密度。鉴于目前几乎没有或不存在定量表征工程构建体的非破坏性检测方法，这种非侵入性方法可能成为符合监管要求的、满足特定质量标准的工程移植物制造中的一种相关工具。
