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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

Bright field and fluorescence images of ABCG2 protein expression level of both type 1 MHCC 97L(a) and type 2 MHCC 97L (b) Scale bars in Figs 7a,b are both 100 μm.
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f7: Bright field and fluorescence images of ABCG2 protein expression level of both type 1 MHCC 97L(a) and type 2 MHCC 97L (b) Scale bars in Figs 7a,b are both 100 μm.

Mentions: Protein expression level is also one aspect of the cell heterogeneity. As we known, ABCG2 protein attached to ABC protein family was commonly expressed in carcinoma cell. Immunofluorescence of ABCG2 protein expression was conducted as showed in Fig. 7. The results confirmed that obvious differences of the protein expression level existed between the two clones. As shown in Figs 7a,b and Fig. S10, the ABCG2 expression level of type 1 MHCC 97L is much higher than that of type 2 MHCC 97L, which further confirmed that heterogeneity existed between the two clones.


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)

Bright field and fluorescence images of ABCG2 protein expression level of both type 1 MHCC 97L(a) and type 2 MHCC 97L (b) Scale bars in Figs 7a,b are both 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f7: Bright field and fluorescence images of ABCG2 protein expression level of both type 1 MHCC 97L(a) and type 2 MHCC 97L (b) Scale bars in Figs 7a,b are both 100 μm.
Mentions: Protein expression level is also one aspect of the cell heterogeneity. As we known, ABCG2 protein attached to ABC protein family was commonly expressed in carcinoma cell. Immunofluorescence of ABCG2 protein expression was conducted as showed in Fig. 7. The results confirmed that obvious differences of the protein expression level existed between the two clones. As shown in Figs 7a,b and Fig. S10, the ABCG2 expression level of type 1 MHCC 97L is much higher than that of type 2 MHCC 97L, which further confirmed that heterogeneity existed between the two clones.

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