Real-time label-free monitoring of adipose-derived stem cell differentiation with electric cell-substrate impedance sensing

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Abstract

Real-time monitoring of stem cells (SCs) differentiation will be critical to scale-up SC technologies, while label-free techniques will be desirable to quality-control SCs without precluding their therapeutic potential. We cultured adipose-derived stem cells (ADSCs) on top of multielectrode arrays and measured variations in the complex impedance Z* throughout induction of ADSCs toward osteoblasts and adipocytes. Z* was measured up to 17 d, every 180 s, over a 62.5-64kHz frequency range with an ECIS Zθ instrument. We found that osteogenesis and adipogenesis were characterized by distinct Z* time-courses. Significant differences were found (P = 0.007) as soon as 12 h post induction. An increase in the barrier resistance (Rb) up to 1.7 ohm·cm2 was associated with early osteo-induction, whereas Rb peaked at 0.63 ohm·cm2 for adipo-induced cells before falling to zero at t = 129 h. Dissimilarities in Z* throughout early induction (<24 h) were essentially attributed to variations in the cell-substrate parameter α. Four days after induction, cell membrane capacitance (Cm) of osteoinduced cells (Cm = 1.72 ± 0.10 μF/cm2) was significantly different from that of adipo-induced cells (Cm = 2.25 ± 0.27 μF/cm2), indicating that Cm could be used as an early marker of differentiation. Finally, we demonstrated long-term monitoring and measured a shift in the complex plane in the middle frequency range (1 kHz to 8 kHz) between early (t = 100 h) and late induction (t = 380 h). This study demonstrated that the osteoblast and adipocyte lineages have distinct dielectric properties and that such differences can be used to perform real-time label-free quantitative monitoring of adult stem cell differentiation with impedance sensing.

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Bagnaninchi, P. O., & Drummond, N. (2011). Real-time label-free monitoring of adipose-derived stem cell differentiation with electric cell-substrate impedance sensing. Proceedings of the National Academy of Sciences of the United States of America, 108(16), 6462–6467. https://doi.org/10.1073/pnas.1018260108

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