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Real-time monitoring of neural differentiation of human mesenchymal stem cells by electric cell-substrate impedance sensing.

Park HE, Kim D, Koh HS, Cho S, Sung JS, Kim JY - J. Biomed. Biotechnol. (2011)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Science, Gachon University of Medicine and Science, Incheon 406-799, Republic of Korea.

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.

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Neuron-specific gene expression of hMSCs during neural differentiation. (a) Cells that had been harvested 6 hr for Nurr-1, 12 hr for NeuroD6, and 48 hr for NF-L and NF-M were used in RT-PCR assay. NGM: normal growth media; NIM: neural induction media; NDM: neural differentiation media. (b) The relative expression of Nurr-1, NF-L, and NF-M was normalized by GAPDH using densitometric analysis. Data for NeuroD6 were omitted because bands corresponding to NeuroD6 were too faint to be measured by densitometry.
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fig3: Neuron-specific gene expression of hMSCs during neural differentiation. (a) Cells that had been harvested 6 hr for Nurr-1, 12 hr for NeuroD6, and 48 hr for NF-L and NF-M were used in RT-PCR assay. NGM: normal growth media; NIM: neural induction media; NDM: neural differentiation media. (b) The relative expression of Nurr-1, NF-L, and NF-M was normalized by GAPDH using densitometric analysis. Data for NeuroD6 were omitted because bands corresponding to NeuroD6 were too faint to be measured by densitometry.

Mentions: Since the numbers of cells expressing the neuron-specific protein marker Tuj-1 in NDM were found to be twice as high as those in the other media, cellular mRNA expressions of several neuron-specific genes were examined by RT-PCR. As shown in Figure 3, the expression levels of neuronal markers such as Nurr-1, NF-L, and NF-M in the NDM group were relatively higher than in the other groups. Overall results indicate that hMSCs cultured in NDM were effectively induced to differentiate into neuronal cells.


Real-time monitoring of neural differentiation of human mesenchymal stem cells by electric cell-substrate impedance sensing.

Park HE, Kim D, Koh HS, Cho S, Sung JS, Kim JY - J. Biomed. Biotechnol. (2011)

Neuron-specific gene expression of hMSCs during neural differentiation. (a) Cells that had been harvested 6 hr for Nurr-1, 12 hr for NeuroD6, and 48 hr for NF-L and NF-M were used in RT-PCR assay. NGM: normal growth media; NIM: neural induction media; NDM: neural differentiation media. (b) The relative expression of Nurr-1, NF-L, and NF-M was normalized by GAPDH using densitometric analysis. Data for NeuroD6 were omitted because bands corresponding to NeuroD6 were too faint to be measured by densitometry.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3116536&req=5

fig3: Neuron-specific gene expression of hMSCs during neural differentiation. (a) Cells that had been harvested 6 hr for Nurr-1, 12 hr for NeuroD6, and 48 hr for NF-L and NF-M were used in RT-PCR assay. NGM: normal growth media; NIM: neural induction media; NDM: neural differentiation media. (b) The relative expression of Nurr-1, NF-L, and NF-M was normalized by GAPDH using densitometric analysis. Data for NeuroD6 were omitted because bands corresponding to NeuroD6 were too faint to be measured by densitometry.
Mentions: Since the numbers of cells expressing the neuron-specific protein marker Tuj-1 in NDM were found to be twice as high as those in the other media, cellular mRNA expressions of several neuron-specific genes were examined by RT-PCR. As shown in Figure 3, the expression levels of neuronal markers such as Nurr-1, NF-L, and NF-M in the NDM group were relatively higher than in the other groups. Overall results indicate that hMSCs cultured in NDM were effectively induced to differentiate into neuronal cells.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Science, Gachon University of Medicine and Science, Incheon 406-799, Republic of Korea.

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.

Show MeSH
Related in: MedlinePlus