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Electrical Property Characterization of Neural Stem Cells in Differentiation.

Zhao Y, Liu Q, Sun H, Chen D, Li Z, Fan B, George J, Xue C, Cui Z, Wang J, Chen J - PLoS ONE (2016)

Bottom Line: In this paper, a microfluidic platform enabling the high-throughput quantification of Cspecific membrane and σcytoplasm from hundreds of single neural stem cells undergoing differentiation was developed to explore the feasibility to characterize the neural stem cell differentiation process without biochemical staining.The recorded electrical properties of neural stem cells undergoing differentiation showed distinctive and unique patterns: 1) in the suspension culture before inducing differentiation, a large distribution and difference in σcytoplasm among individual neural stem cells was noticed, which indicated heterogeneity that may result from the nature of suspension culture of neurospheres; and 2) during the differentiation in adhering monolayer culture, significant changes and a large difference in Cspecific membrane were located indicating different expressions of membrane proteins during the differentiation process, and a small distribution difference in σcytoplasm was less significant that indicated the relatively consistent properties of cytoplasm during the culture.In summary, significant differences in Cspecific membrane and σcytoplasm were observed during the neural stem cell differentiation process, which may potentially be used as label-free biophysical markers to monitor this process.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing, P.R. China.

ABSTRACT
Electrical property characterization of stem cells could be utilized as a potential label-free biophysical approach to evaluate the differentiation process. However, there has been a lack of technology or tools that can quantify the intrinsic cellular electrical markers (e.g., specific membrane capacitance (Cspecific membrane) and cytoplasm conductivity (σcytoplasm)) for a large amount of stem cells or differentiated cells. In this paper, a microfluidic platform enabling the high-throughput quantification of Cspecific membrane and σcytoplasm from hundreds of single neural stem cells undergoing differentiation was developed to explore the feasibility to characterize the neural stem cell differentiation process without biochemical staining. Experimental quantification using biochemical markers (e.g., Nestin, Tubulin and GFAP) of neural stem cells confirmed the initiation of the differentiation process featured with gradual loss in cellular stemness and increased cell markers for neurons and glial cells. The recorded electrical properties of neural stem cells undergoing differentiation showed distinctive and unique patterns: 1) in the suspension culture before inducing differentiation, a large distribution and difference in σcytoplasm among individual neural stem cells was noticed, which indicated heterogeneity that may result from the nature of suspension culture of neurospheres; and 2) during the differentiation in adhering monolayer culture, significant changes and a large difference in Cspecific membrane were located indicating different expressions of membrane proteins during the differentiation process, and a small distribution difference in σcytoplasm was less significant that indicated the relatively consistent properties of cytoplasm during the culture. In summary, significant differences in Cspecific membrane and σcytoplasm were observed during the neural stem cell differentiation process, which may potentially be used as label-free biophysical markers to monitor this process.

No MeSH data available.


Related in: MedlinePlus

Typical impedance amplitude and phase measurement at 1 kHz and 100 kHz, simultaneously as well as cellular images squeezing through the constriction channels for a single neural stem cell for rat 1 on Day 1.During the cell squeezing process, there is an amplitude increase and phase decrease for data at both 1 kHz and 100 kHz. Basal impedance amplitudes (no cell entry) at 1 kHz were lower than the values at 100 kHz and there were higher impedance amplitude increases during the cellular squeezing process at 1 kHz than those of 100 kHz. As to the phase data, basal phase values (no cell entry) at 1 kHz were almost zero degree, which were higher than the values at 100 kHz. There were higher phase changes during the cellular squeezing process at 100 kHz than 1 kHz.
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pone.0158044.g003: Typical impedance amplitude and phase measurement at 1 kHz and 100 kHz, simultaneously as well as cellular images squeezing through the constriction channels for a single neural stem cell for rat 1 on Day 1.During the cell squeezing process, there is an amplitude increase and phase decrease for data at both 1 kHz and 100 kHz. Basal impedance amplitudes (no cell entry) at 1 kHz were lower than the values at 100 kHz and there were higher impedance amplitude increases during the cellular squeezing process at 1 kHz than those of 100 kHz. As to the phase data, basal phase values (no cell entry) at 1 kHz were almost zero degree, which were higher than the values at 100 kHz. There were higher phase changes during the cellular squeezing process at 100 kHz than 1 kHz.

Mentions: Typical impedance amplitude and phase measurement at 1 kHz and 100 kHz, simultaneously as well as images of cellular squeezing through the constriction channels for a neural stem cells of rat 1 on Day 1 were shown in Fig 3. During the cell squeezing process, there was an amplitude increase and a phase decrease for impedance data at both 1 kHz and 100 kHz. Basal impedance amplitudes (no cell entry) at 1 kHz were lower than the values at 100 kHz and there were higher impedance amplitude increases during the cellular squeezing process at 1 kHz than those of 100 kHz. As to the phase data, basal phase values (no cell entry) at 1 kHz were almost zero degree, which were higher than the values at 100 kHz. There were higher phase changes during the cellular squeezing process at 100 kHz than 1 kHz. The similar trend was observed from other testing time point and another rat as well.


Electrical Property Characterization of Neural Stem Cells in Differentiation.

Zhao Y, Liu Q, Sun H, Chen D, Li Z, Fan B, George J, Xue C, Cui Z, Wang J, Chen J - PLoS ONE (2016)

Typical impedance amplitude and phase measurement at 1 kHz and 100 kHz, simultaneously as well as cellular images squeezing through the constriction channels for a single neural stem cell for rat 1 on Day 1.During the cell squeezing process, there is an amplitude increase and phase decrease for data at both 1 kHz and 100 kHz. Basal impedance amplitudes (no cell entry) at 1 kHz were lower than the values at 100 kHz and there were higher impedance amplitude increases during the cellular squeezing process at 1 kHz than those of 100 kHz. As to the phase data, basal phase values (no cell entry) at 1 kHz were almost zero degree, which were higher than the values at 100 kHz. There were higher phase changes during the cellular squeezing process at 100 kHz than 1 kHz.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4920408&req=5

pone.0158044.g003: Typical impedance amplitude and phase measurement at 1 kHz and 100 kHz, simultaneously as well as cellular images squeezing through the constriction channels for a single neural stem cell for rat 1 on Day 1.During the cell squeezing process, there is an amplitude increase and phase decrease for data at both 1 kHz and 100 kHz. Basal impedance amplitudes (no cell entry) at 1 kHz were lower than the values at 100 kHz and there were higher impedance amplitude increases during the cellular squeezing process at 1 kHz than those of 100 kHz. As to the phase data, basal phase values (no cell entry) at 1 kHz were almost zero degree, which were higher than the values at 100 kHz. There were higher phase changes during the cellular squeezing process at 100 kHz than 1 kHz.
Mentions: Typical impedance amplitude and phase measurement at 1 kHz and 100 kHz, simultaneously as well as images of cellular squeezing through the constriction channels for a neural stem cells of rat 1 on Day 1 were shown in Fig 3. During the cell squeezing process, there was an amplitude increase and a phase decrease for impedance data at both 1 kHz and 100 kHz. Basal impedance amplitudes (no cell entry) at 1 kHz were lower than the values at 100 kHz and there were higher impedance amplitude increases during the cellular squeezing process at 1 kHz than those of 100 kHz. As to the phase data, basal phase values (no cell entry) at 1 kHz were almost zero degree, which were higher than the values at 100 kHz. There were higher phase changes during the cellular squeezing process at 100 kHz than 1 kHz. The similar trend was observed from other testing time point and another rat as well.

Bottom Line: In this paper, a microfluidic platform enabling the high-throughput quantification of Cspecific membrane and σcytoplasm from hundreds of single neural stem cells undergoing differentiation was developed to explore the feasibility to characterize the neural stem cell differentiation process without biochemical staining.The recorded electrical properties of neural stem cells undergoing differentiation showed distinctive and unique patterns: 1) in the suspension culture before inducing differentiation, a large distribution and difference in σcytoplasm among individual neural stem cells was noticed, which indicated heterogeneity that may result from the nature of suspension culture of neurospheres; and 2) during the differentiation in adhering monolayer culture, significant changes and a large difference in Cspecific membrane were located indicating different expressions of membrane proteins during the differentiation process, and a small distribution difference in σcytoplasm was less significant that indicated the relatively consistent properties of cytoplasm during the culture.In summary, significant differences in Cspecific membrane and σcytoplasm were observed during the neural stem cell differentiation process, which may potentially be used as label-free biophysical markers to monitor this process.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing, P.R. China.

ABSTRACT
Electrical property characterization of stem cells could be utilized as a potential label-free biophysical approach to evaluate the differentiation process. However, there has been a lack of technology or tools that can quantify the intrinsic cellular electrical markers (e.g., specific membrane capacitance (Cspecific membrane) and cytoplasm conductivity (σcytoplasm)) for a large amount of stem cells or differentiated cells. In this paper, a microfluidic platform enabling the high-throughput quantification of Cspecific membrane and σcytoplasm from hundreds of single neural stem cells undergoing differentiation was developed to explore the feasibility to characterize the neural stem cell differentiation process without biochemical staining. Experimental quantification using biochemical markers (e.g., Nestin, Tubulin and GFAP) of neural stem cells confirmed the initiation of the differentiation process featured with gradual loss in cellular stemness and increased cell markers for neurons and glial cells. The recorded electrical properties of neural stem cells undergoing differentiation showed distinctive and unique patterns: 1) in the suspension culture before inducing differentiation, a large distribution and difference in σcytoplasm among individual neural stem cells was noticed, which indicated heterogeneity that may result from the nature of suspension culture of neurospheres; and 2) during the differentiation in adhering monolayer culture, significant changes and a large difference in Cspecific membrane were located indicating different expressions of membrane proteins during the differentiation process, and a small distribution difference in σcytoplasm was less significant that indicated the relatively consistent properties of cytoplasm during the culture. In summary, significant differences in Cspecific membrane and σcytoplasm were observed during the neural stem cell differentiation process, which may potentially be used as label-free biophysical markers to monitor this process.

No MeSH data available.


Related in: MedlinePlus