Diversification of Primary Mesenchymal Stromal/Stem Cell Populations at the Single-Cell Level Using Raman Spectroscopy

Abstract

Abstract. As a promising optical technique for application in biomedicine, Raman spectroscopy has been used for stem cell analysis, whereby the largest number of studies was based on the examination of the differentiation status of mesenchymal stromal/stem cells (MSCs) [1,2]. Namely, MSCs represent a diverse population of multipotent precursors that reside in many tissues. They have been isolated from various tissues and organs including bone marrow, adipose tissue, teeth, amniotic fluid, umbilical cord, tendon, etc., and due to simple and non invasive isolation procedures MSCs are considered a valuable alternative source for cell replacement therapies. The main features of these cells are the ability to self-renew and the differentiation into several types of mature cells such as osteoblasts, adipocytes, chondrocytes under in vitro conditions. However, there is no precise marker that can be used to isolate and characterize this cell population, which significantly hinders further progress in potential application of these cells for therapeutic purposes. Therefore, our goal was to investigate the use of Raman spectroscopy to characterize biochemical profile of MSCs at single-cell level. In this study primary human MSCs derived from bone marrow (BM-MSCs) of five healthy pediatric donors collected during allogenic transplantation were analyzed. By using standard biological tests related to the MSCs features such as adherence, phenotype, clonogenicity, proliferation rate, pluripotency and multilineage differentiation potential variations between these donors were not detected. Raman spectroscopy analysis of MSCs at the single-cell level revealed a similar biochemical background of the tested samples. However, following the extensive principal component analysis (PCA), a clustering of MSCs populations was detected, particularly when the samples were analyzed in pairs. Obtained results indicate that Raman spectroscopy technique could provide valuable information for MSCs diversification and contribute to MSCs characterization, consequently accelerating their application in cell therap