Stem cell studies hold enormous potential for development of new therapies with unique differentiation ability of stem cells into various cell types. In this manner, investigation of stem cells in normal developmental and physiological states as well as in pathological conditions may lead to understanding of disease pathogenesis and development of new cellular therapies. The chemical changes responsible for a stem cell's unique physiological properties have been explored by various invasive techniques. The biological techniques being used for stem cell analyses involve fixing, staining, and chemical drying. All these steps have damaging effect on life processes of stem cells and they do not provide real-time information about cellular conditions. Therefore; newly non-destructive research methods that enable real-time chemical monitoring, high-quality data collection with less experimental complexity and expense must be used for stem cell characterization. In this context, vibrational spectroscopic appoaches can provide promissing alternatives to get new information into the field of stem cell biology, and they can be successfully used for chemical analysis, quantification and imaging of stem cells. The application of new complimentary vibrational spectroscopic aproaches will shed light on stem cell biology and will provide new insigth into stem cell reseaches for future therapeutic and regenerative medicines.