Plant suspension culture is attracting interest as a promising platform to produce biological medicines due to the absence of virus, prions or DNA related to mammals during the production process. However, the heterogenic plant cell proliferation nature is particularly challenging for establishing industrial processes based on innovative approaches currently used, particularly in the animal cell culture industry. In this context, while Process Analytical Technology (PAT) tools have been used to monitor classical parameters such as biomass dry weight, its use in cells heterogeneity has received limited attention. Therefore, the feasibility of in situ monitoring of cell differentiation in plant cell suspensions employing NIR spectroscopy and chemometrics was investigated. Off-line measurements of cell heterogeneity in term of cell differentiation and in-line NIR spectra captured in 3 L bioreactor cultures were employed to generate calibration models. Then models were tested to estimate the population distribution of parenchyma, collenchyma and sclerenchyma cells during Catharanthus roseus suspension cultures. Results have proven in situ NIR spectroscopy as a capable PAT tool to monitor differentiated cells accurately and in real-time. These results are the starting point to follow-up PAT systems so that plant cell culture heterogeneity may be better understood and controlled in biopharmaceutical plant cell cultures.

中文翻译：

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通过近红外光谱技术对长春花悬浮培养过程进行原位细胞分化监测。

由于在生产过程中不存在与哺乳动物有关的病毒，病毒或DNA，因此植物悬浮培养作为生产生物药物的有前途的平台引起了人们的兴趣。然而，异源植物细胞增殖的性质对于基于当前使用的创新方法建立工业过程尤其具有挑战性，特别是在动物细胞培养工业中。在这种情况下，虽然过程分析技术（PAT）工具已用于监视经典参数，例如生物质干重，但其在细胞异质性中的应用受到了有限的关注。因此，研究了利用NIR光谱学和化学计量学原位监测植物细胞悬液中细胞分化的可行性。根据细胞分化和在3 L生物反应器培养物中捕获的在线近红外光谱对细胞异质性进行离线测量，以生成校准模型。然后测试模型以估计长春花悬浮培养期间薄壁组织，间质细胞和硬化细胞的分布。结果证明了原位近红外光谱技术是一种功能强大的PAT工具，可以准确，实时地监测分化的细胞。这些结果是后续PAT系统的起点，因此可以在生物制药植物细胞培养物中更好地理解和控制植物细胞培养的异质性。长春花悬浮培养过程中的肾盂和巩膜细胞。结果证明了原位近红外光谱技术是一种功能强大的PAT工具，可以准确，实时地监测分化的细胞。这些结果是后续PAT系统的起点，因此可以在生物制药植物细胞培养物中更好地理解和控制植物细胞培养的异质性。长春花悬浮培养中的红细胞和硬化细胞。结果证明了原位近红外光谱技术是一种功能强大的PAT工具，可以准确，实时地监测分化的细胞。这些结果是后续PAT系统的起点，因此可以在生物制药植物细胞培养物中更好地理解和控制植物细胞培养的异质性。