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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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Morphological and flow cytometric characterization of primary human mesenchymal stem cells (hMSCs). (a) Morphology of primary human umbilical cord blood-derived MSC cultured in normal growth medium. hMSCs at passages 8-9 have a fibroblast-like morphology. ×100. (b) Immunophenotyping results of human umbilical cord blood-derived MSCs. Cells at passages 8-9 were stained with antibodies listed in Section 2 and assayed by flow cytometry. The cells were strongly positive for MSC-specific markers, such as CD13, CD44, CD73, CD90, and CD105, while negative for the hematopoietic stem cell markers CD14, CD34, and CD45. Staining for nonspecific immunoglobulin G (IgG) isotype fluorescence used as a control is represented by gray fill.
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fig1: Morphological and flow cytometric characterization of primary human mesenchymal stem cells (hMSCs). (a) Morphology of primary human umbilical cord blood-derived MSC cultured in normal growth medium. hMSCs at passages 8-9 have a fibroblast-like morphology. ×100. (b) Immunophenotyping results of human umbilical cord blood-derived MSCs. Cells at passages 8-9 were stained with antibodies listed in Section 2 and assayed by flow cytometry. The cells were strongly positive for MSC-specific markers, such as CD13, CD44, CD73, CD90, and CD105, while negative for the hematopoietic stem cell markers CD14, CD34, and CD45. Staining for nonspecific immunoglobulin G (IgG) isotype fluorescence used as a control is represented by gray fill.

Mentions: To determine whether umbilical cord-derived hMSCs purchased from Promocell had MSC characteristics, we examined the cells for morphologic characteristics and expression of MSC-specific markers. Figure 1(a) shows that primary MSCs display a typical fibroblast-like morphology. These cells expressed MSC specific markers such as CD13, CD44, CD105, CD90, and CD73 but not hematopoietic stem cell markers such as CD14, CD34, and CD45 (Figure 1(b)).


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)

Morphological and flow cytometric characterization of primary human mesenchymal stem cells (hMSCs). (a) Morphology of primary human umbilical cord blood-derived MSC cultured in normal growth medium. hMSCs at passages 8-9 have a fibroblast-like morphology. ×100. (b) Immunophenotyping results of human umbilical cord blood-derived MSCs. Cells at passages 8-9 were stained with antibodies listed in Section 2 and assayed by flow cytometry. The cells were strongly positive for MSC-specific markers, such as CD13, CD44, CD73, CD90, and CD105, while negative for the hematopoietic stem cell markers CD14, CD34, and CD45. Staining for nonspecific immunoglobulin G (IgG) isotype fluorescence used as a control is represented by gray fill.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig1: Morphological and flow cytometric characterization of primary human mesenchymal stem cells (hMSCs). (a) Morphology of primary human umbilical cord blood-derived MSC cultured in normal growth medium. hMSCs at passages 8-9 have a fibroblast-like morphology. ×100. (b) Immunophenotyping results of human umbilical cord blood-derived MSCs. Cells at passages 8-9 were stained with antibodies listed in Section 2 and assayed by flow cytometry. The cells were strongly positive for MSC-specific markers, such as CD13, CD44, CD73, CD90, and CD105, while negative for the hematopoietic stem cell markers CD14, CD34, and CD45. Staining for nonspecific immunoglobulin G (IgG) isotype fluorescence used as a control is represented by gray fill.
Mentions: To determine whether umbilical cord-derived hMSCs purchased from Promocell had MSC characteristics, we examined the cells for morphologic characteristics and expression of MSC-specific markers. Figure 1(a) shows that primary MSCs display a typical fibroblast-like morphology. These cells expressed MSC specific markers such as CD13, CD44, CD105, CD90, and CD73 but not hematopoietic stem cell markers such as CD14, CD34, and CD45 (Figure 1(b)).

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