SERS-based long-term mitochondrial pH monitoring during differentiation of human induced pluripotent stem cells to neural progenitor cells.

As one of the important organelles in the process of cell differentiation, mitochondria regulate the whole process of differentiation by participating in energy supply and information transmission. Mitochondrial pH value is a key indicator of mitochondrial function. Therefore, real-time monitoring of mitochondrial pH value during cell differentiation is of great significance for understanding cell biochemical processes and exploring differentiation mechanisms. In this study, Surface-enhanced Raman scattering (SERS) technology was used to achieve the real-time monitoring of mitochondrial pH during induced pluripotent stem cells (iPSCs) differentiation into neural progenitor cells (NPCs). The results showed that the variation trend of mitochondrial pH in normal and abnormal differentiated batches was different. The mitochondrial pH value of normal differentiated cells continued to decline from iPSCs to embryoid bodies (EB) day 4, and continued to rise from EB day 4 to the NPCs stage, and the mitochondrial microenvironment of iPSCs to NPCs differentiation became acidic. In contrast, the mitochondrial pH value of abnormally differentiated cells declined continuously during differentiation. This study improves the information on acid-base balance during cell differentiation and may provide a basis for further understanding of the changes and regulatory mechanisms of mitochondrial metabolism during cell differentiation. This also helps to improve more accurate and useful differentiation protocols based on the microenvironment within the mitochondria, improving the efficiency of cell differentiation.