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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

(a) Schematic of the experimental design composed of stem cell isolation and differentiation with biochemical and bioelectrical parameters quantified. The cell culture steps include isolation of neural stem cells from rats, suspension culture of neural stem cells (Day 0) and differentiation of neural stems in the adhesive culture (Day 1, Day 3, and Day 7). On Day 0, Day 1, Day 3 and Day 7, two portions of neural stem cells were collected for the characterization of cellular biochemical parameters based on RT-PCR and bioelectrical parameters based on the home-developed impedance microfluidic flow cytometry, respectively. (b) Experimental setup of the microfluidic system for continuous characterization of Cspecific membrane and σcytoplasm of single cells in suspension where cells are aspirated continuously through the constriction channel with impedance data at 1 kHz and 100 kHz and cell elongation length measured by a lock-in amplifier and an inverted microscope simultaneously.
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pone.0158044.g001: (a) Schematic of the experimental design composed of stem cell isolation and differentiation with biochemical and bioelectrical parameters quantified. The cell culture steps include isolation of neural stem cells from rats, suspension culture of neural stem cells (Day 0) and differentiation of neural stems in the adhesive culture (Day 1, Day 3, and Day 7). On Day 0, Day 1, Day 3 and Day 7, two portions of neural stem cells were collected for the characterization of cellular biochemical parameters based on RT-PCR and bioelectrical parameters based on the home-developed impedance microfluidic flow cytometry, respectively. (b) Experimental setup of the microfluidic system for continuous characterization of Cspecific membrane and σcytoplasm of single cells in suspension where cells are aspirated continuously through the constriction channel with impedance data at 1 kHz and 100 kHz and cell elongation length measured by a lock-in amplifier and an inverted microscope simultaneously.

Mentions: The experimental design and bioelectrical characterisation set-up were shown in Fig 1. Neural stem cells, initially cultured in the suspension state (Day 0), were transferred to adhesive culture to start the differentiation process (Day 1, Day 3 and Day 7). On Day 0, Day 1, Day 3 and Day 7, two portions of neural stem cells were collected for the characterization of cellular biochemical markers (e.g., Nestin, Tubulin and GFAP) based on RT-PCR and bioelectrical parameters (e.g., Cspecific membrane and σcytoplasm) based on the microfluidic platform, respectively.


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)

(a) Schematic of the experimental design composed of stem cell isolation and differentiation with biochemical and bioelectrical parameters quantified. The cell culture steps include isolation of neural stem cells from rats, suspension culture of neural stem cells (Day 0) and differentiation of neural stems in the adhesive culture (Day 1, Day 3, and Day 7). On Day 0, Day 1, Day 3 and Day 7, two portions of neural stem cells were collected for the characterization of cellular biochemical parameters based on RT-PCR and bioelectrical parameters based on the home-developed impedance microfluidic flow cytometry, respectively. (b) Experimental setup of the microfluidic system for continuous characterization of Cspecific membrane and σcytoplasm of single cells in suspension where cells are aspirated continuously through the constriction channel with impedance data at 1 kHz and 100 kHz and cell elongation length measured by a lock-in amplifier and an inverted microscope simultaneously.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0158044.g001: (a) Schematic of the experimental design composed of stem cell isolation and differentiation with biochemical and bioelectrical parameters quantified. The cell culture steps include isolation of neural stem cells from rats, suspension culture of neural stem cells (Day 0) and differentiation of neural stems in the adhesive culture (Day 1, Day 3, and Day 7). On Day 0, Day 1, Day 3 and Day 7, two portions of neural stem cells were collected for the characterization of cellular biochemical parameters based on RT-PCR and bioelectrical parameters based on the home-developed impedance microfluidic flow cytometry, respectively. (b) Experimental setup of the microfluidic system for continuous characterization of Cspecific membrane and σcytoplasm of single cells in suspension where cells are aspirated continuously through the constriction channel with impedance data at 1 kHz and 100 kHz and cell elongation length measured by a lock-in amplifier and an inverted microscope simultaneously.
Mentions: The experimental design and bioelectrical characterisation set-up were shown in Fig 1. Neural stem cells, initially cultured in the suspension state (Day 0), were transferred to adhesive culture to start the differentiation process (Day 1, Day 3 and Day 7). On Day 0, Day 1, Day 3 and Day 7, two portions of neural stem cells were collected for the characterization of cellular biochemical markers (e.g., Nestin, Tubulin and GFAP) based on RT-PCR and bioelectrical parameters (e.g., Cspecific membrane and σcytoplasm) based on the microfluidic platform, respectively.

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