Abstract
Stem cells are useful for cell replacement therapy. Stem cell differentiation must be monitored thoroughly and precisely prior to transplantation. In this study we evaluated the usefulness of electric cell-substrate impedance sensing (ECIS) for in vitro real-time monitoring of neural differentiation of human mesenchymal stem cells (hMSCs). We cultured hMSCs in neural differentiation media (NDM) for 6 days and examined the time-course of impedance changes with an ECIS array. We also monitored the expression of markers for neural differentiation, total cell count, and cell cycle profiles. Cellular expression of neuron and oligodendrocyte markers increased. The resistance value of cells cultured in NDM was automatically measured in real-time and found to increase much more slowly over time compared to cells cultured in non-differentiation media. The relatively slow resistance changes observed in differentiating MSCs were determined to be due to their lower growth capacity achieved by induction of cell cycle arrest in G0/G1. Overall results suggest that the relatively slow change in resistance values measured by ECIS method can be used as a parameter for slowly growing neural-differentiating cells. However, to enhance the competence of ECIS for in vitro real-time monitoring of neural differentiation of MSCs, more elaborate studies are needed.
Highlights
Due to their long-term self-renewal capacity and multilineage differentiation potential, stem cells have been considered as useful replacement material to heal cellular injuries caused by trauma, infection, and genetic diseases
To determine whether umbilical cord-derived human mesenchymal stem cells (hMSCs) purchased from Promocell had MSC characteristics, we examined the cells for morphologic characteristics and expression of MSC-specific markers
We examined the resistance values of the cell layer in 3 different culture media throughout the culture period to determine whether electric cell-substrate impedance sensing (ECIS) is useful for real-time monitoring of neural differentiation of hMSCs
Summary
Due to their long-term self-renewal capacity and multilineage differentiation potential, stem cells have been considered as useful replacement material to heal cellular injuries caused by trauma, infection, and genetic diseases. The purity and yield of differentiated cells are critical for successful stem cell therapy [1]. For these reasons, monitoring the process of in vitro stem cell differentiation is important. All the methods mentioned are labor-intensive multistep processes, which are end-point assays that offer only a snapshot of what is occurring. These techniques usually involve labeling with nucleic acids or antibodies and destruction of the cells. We performed neural differentiation of human umbilical cord-derived mesenchymal stem cells (hMSCs) according to a previously established differentiation protocol [12] and attempted to evaluate the usefulness of ECIS for real-time monitoring of differentiation by measuring the resistance change of the cell layer
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