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A High Throughput Micro-Chamber Array Device for Single Cell Clonal Cultivation and Tumor Heterogeneity Analysis.

Shen FM, Zhu L, Ye H, Yang YJ, Pang DW, Zhang ZL - Sci Rep (2015)

Bottom Line: Recently, single cell cloning techniques have been gradually developed benefited from their important roles in monoclonal antibody screening, tumor heterogeneity research fields, etc.In this study, we developed a high throughput device containing 1400 lateral chambers to efficiently isolate single cells and carry out long-term single cell clonal cultivation as well as tumor heterogeneity studies.The device was further used to examine tumor heterogeneity such as morphology, growth rate, anti-cancer drug tolerance as well as adenosine triphosphate-binding cassette (ABC) transporter ABCG2 protein expression level.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, P. R. China.

ABSTRACT
Recently, single cell cloning techniques have been gradually developed benefited from their important roles in monoclonal antibody screening, tumor heterogeneity research fields, etc. In this study, we developed a high throughput device containing 1400 lateral chambers to efficiently isolate single cells and carry out long-term single cell clonal cultivation as well as tumor heterogeneity studies. Most of the isolated single cells could proliferate normally nearly as long as three weeks and hundreds of clones could be formed once with one device, which made it possible to study tumor heterogeneity at single cell level. The device was further used to examine tumor heterogeneity such as morphology, growth rate, anti-cancer drug tolerance as well as adenosine triphosphate-binding cassette (ABC) transporter ABCG2 protein expression level. Except for the single cell isolation and tumor heterogeneity studies, the device is expected to be used as an excellent platform for drug screening, tumor biomarker discovering and tumor metastasis assay.

No MeSH data available.


Related in: MedlinePlus

Fluorescence images of cell apoptosis under incubation with indomethacin.(a) type 1 MHCC 97L apoptosis images under the effect of indomethacin. (b) type 2 MHCC 97L apoptosis images under the effect of indomethacin. (c), (d) fluorescence images of type 1 MHCC 97L and type 2 MHCC 97L apoptosis images in control group. The red, green and blue fluorescence come from PI, FITC and Hoechst 33342, respectively. Scale bars in Figs 6a–d are all 100 μm.
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f6: Fluorescence images of cell apoptosis under incubation with indomethacin.(a) type 1 MHCC 97L apoptosis images under the effect of indomethacin. (b) type 2 MHCC 97L apoptosis images under the effect of indomethacin. (c), (d) fluorescence images of type 1 MHCC 97L and type 2 MHCC 97L apoptosis images in control group. The red, green and blue fluorescence come from PI, FITC and Hoechst 33342, respectively. Scale bars in Figs 6a–d are all 100 μm.

Mentions: According to recent report41, indomethacin inhibits hepatoma carcinoma cell proliferation. We here chose 0.3 mmol/L indomethacin (according to the half maximal inhibitory concentration namely IC50 experiment, Fig. S8) to examine its cytotoxicity on the two clones. An AnnexinV-FITC/PI apoptosis kit was used to confirm the cytotoxicity of indomethacin to the clones. The red and green fluorescence were respectively emitted from propidium iodide (PI) fluorescein isothiocyanate (FITC) of AnnexinV-FITC/PI apoptosis kit. Cells labeled with red fluorescence or green fluorescence were considered as apoptotic cells. Compared with the weak fluorescence intensity displayed in the group of type 2 MHCC 97L (Fig. 6b), apparent apoptosis phenomenon appeared among type 1 MHCC 97L as shown in Fig. 6a.


A High Throughput Micro-Chamber Array Device for Single Cell Clonal Cultivation and Tumor Heterogeneity Analysis.

Shen FM, Zhu L, Ye H, Yang YJ, Pang DW, Zhang ZL - Sci Rep (2015)

Fluorescence images of cell apoptosis under incubation with indomethacin.(a) type 1 MHCC 97L apoptosis images under the effect of indomethacin. (b) type 2 MHCC 97L apoptosis images under the effect of indomethacin. (c), (d) fluorescence images of type 1 MHCC 97L and type 2 MHCC 97L apoptosis images in control group. The red, green and blue fluorescence come from PI, FITC and Hoechst 33342, respectively. Scale bars in Figs 6a–d are all 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493670&req=5

f6: Fluorescence images of cell apoptosis under incubation with indomethacin.(a) type 1 MHCC 97L apoptosis images under the effect of indomethacin. (b) type 2 MHCC 97L apoptosis images under the effect of indomethacin. (c), (d) fluorescence images of type 1 MHCC 97L and type 2 MHCC 97L apoptosis images in control group. The red, green and blue fluorescence come from PI, FITC and Hoechst 33342, respectively. Scale bars in Figs 6a–d are all 100 μm.
Mentions: According to recent report41, indomethacin inhibits hepatoma carcinoma cell proliferation. We here chose 0.3 mmol/L indomethacin (according to the half maximal inhibitory concentration namely IC50 experiment, Fig. S8) to examine its cytotoxicity on the two clones. An AnnexinV-FITC/PI apoptosis kit was used to confirm the cytotoxicity of indomethacin to the clones. The red and green fluorescence were respectively emitted from propidium iodide (PI) fluorescein isothiocyanate (FITC) of AnnexinV-FITC/PI apoptosis kit. Cells labeled with red fluorescence or green fluorescence were considered as apoptotic cells. Compared with the weak fluorescence intensity displayed in the group of type 2 MHCC 97L (Fig. 6b), apparent apoptosis phenomenon appeared among type 1 MHCC 97L as shown in Fig. 6a.

Bottom Line: Recently, single cell cloning techniques have been gradually developed benefited from their important roles in monoclonal antibody screening, tumor heterogeneity research fields, etc.In this study, we developed a high throughput device containing 1400 lateral chambers to efficiently isolate single cells and carry out long-term single cell clonal cultivation as well as tumor heterogeneity studies.The device was further used to examine tumor heterogeneity such as morphology, growth rate, anti-cancer drug tolerance as well as adenosine triphosphate-binding cassette (ABC) transporter ABCG2 protein expression level.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, P. R. China.

ABSTRACT
Recently, single cell cloning techniques have been gradually developed benefited from their important roles in monoclonal antibody screening, tumor heterogeneity research fields, etc. In this study, we developed a high throughput device containing 1400 lateral chambers to efficiently isolate single cells and carry out long-term single cell clonal cultivation as well as tumor heterogeneity studies. Most of the isolated single cells could proliferate normally nearly as long as three weeks and hundreds of clones could be formed once with one device, which made it possible to study tumor heterogeneity at single cell level. The device was further used to examine tumor heterogeneity such as morphology, growth rate, anti-cancer drug tolerance as well as adenosine triphosphate-binding cassette (ABC) transporter ABCG2 protein expression level. Except for the single cell isolation and tumor heterogeneity studies, the device is expected to be used as an excellent platform for drug screening, tumor biomarker discovering and tumor metastasis assay.

No MeSH data available.


Related in: MedlinePlus