Martin A. B. Hedegaard

To achieve successful cultivation of pluripotent stem cells using suspension (3D) culture methods, it is essential to maintain the pluripotent state while stimulating the proliferation of the cells. Currently, the methods available to rapidly assess the pluripotency during expansion are limited. Most rely on time‐consuming post hoc ‐analyses, which are prone to operator‐variation, to assess this critical attribute. In this study we propose the use of Raman spectroscopy coupled with chemometric analysis as a process analytical tool to classify the pluripotency by using the spent cultivation medium as samples. Using Support Vector Machines (SVM), a classification accuracy of 94% is achieved on a validation set of various media samples. This study indicates that there may be increased possibilities for monitoring and control in large‐scale expansion of stem cells by Raman spectroscopy.