In situ label-free monitoring of human adipose-derived mesenchymal stem cell differentiation into multiple lineages.

Biomaterials

School of Integrative Engineering, Chung-Ang University, 84 Heukseuk-ro, Dongjak-gu, Seoul 06974, Republic of Korea; Integrative Research Center for Two-Dimensional Functional Materials, Institute of Interdisciplinary Convergence Research, Chung-Ang University, Seoul 06974, Republic of Korea. Electronic address:

Published: February 2018

AI Article Synopsis

  • A new non-invasive method has been developed for quantifying stem cell differentiation into specific cell types at the single-cell level, which is vital for creating patient-specific cells without losing many differentiated cells.
  • This technique uses micro-Raman spectroscopy to analyze human adipose-derived mesenchymal stem cells (hADMSCs) as they differentiate into osteoblasts (bone cells) and adipocytes (fat cells), providing clear and specific signals for each cell type.
  • The method allows for earlier detection of osteogenesis and better visualization of adipogenesis compared to traditional methods, offering significant potential for advancing regenerative therapies.

Article Abstract

Precise characterizations of stem cell differentiation into specific lineages, especially in non-destructive and non-invasive manner, are extremely important for generating patient-specific cells without mass loss of differentiated cells. Here, we report a new method capable of in situ label-free quantification of stem cell differentiation into multiple lineages, even at a single cell level. The human adipose-derived mesenchymal stem cells (hADMSCs) were first differentiated into two different types of cells (osteoblasts and adipocytes) and these differentiated cells were then intensively analyzed by micro-Raman spectroscopy. Interestingly, the Raman peaks assigned to lipid droplets and hydroxyapatite were found to be highly specific to the adipocyte (fat cell) and osteoblast (bone cell) and were thus found to be useful for generating label-free single cell Raman images in combination with CH (2935 cm) peaks for visualizing cell shape. Remarkably, based on these Raman images, we found that the osteogenesis of hADMSCs could be determined and quantified after 9 days of differentiation, which is a week earlier than with the typical alizarin red staining method. In the case of adipogenesis, the increase of lipid droplets in the cytoplasm at the single cell level could be clearly visualized and detected during the entire period of adipogenesis, which is impossible using any other currently available methods such as Oil Red O and immunostaining. Hence, the new method reported in this study is highly promising as an analytical tool for precise in-situ monitoring of stem cell differentiation, and could facilitate the use of stem cell-based materials for the regenerative therapies.

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http://dx.doi.org/10.1016/j.biomaterials.2017.11.005DOI Listing

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