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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

Clones proliferation and morphologies distribution in chambers (days 0–14) starting from a single MHCC 97L cell.(a) type 1 MHCC 97L, b) type 2 MHCC 97L. Scale bars in Figs 4a,b are both 100 μm.
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f4: Clones proliferation and morphologies distribution in chambers (days 0–14) starting from a single MHCC 97L cell.(a) type 1 MHCC 97L, b) type 2 MHCC 97L. Scale bars in Figs 4a,b are both 100 μm.

Mentions: Tumor heterogeneity includes the following aspects namely size difference, different morphologies, relative protein expression level, growth rate and sensitivity to anti-cancer drug and so on3. The text above had displayed that hundreds of clones could be formed in one device once. Among these clones, two apparent cell morphologies were discovered. Most of the clones grew adherently as multilayer and aggregates, we called them type 1 MHCC 97L (Fig. 4a), the other clone grew adherently as a uniform monolayer, and we called them type 2 MHCC 97L (Fig. 4b). Based on the statistics, the percentages of the type 1 and type 2 clones are respectively 97% and 3% (Fig. S7).


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)

Clones proliferation and morphologies distribution in chambers (days 0–14) starting from a single MHCC 97L cell.(a) type 1 MHCC 97L, b) type 2 MHCC 97L. Scale bars in Figs 4a,b are both 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Clones proliferation and morphologies distribution in chambers (days 0–14) starting from a single MHCC 97L cell.(a) type 1 MHCC 97L, b) type 2 MHCC 97L. Scale bars in Figs 4a,b are both 100 μm.
Mentions: Tumor heterogeneity includes the following aspects namely size difference, different morphologies, relative protein expression level, growth rate and sensitivity to anti-cancer drug and so on3. The text above had displayed that hundreds of clones could be formed in one device once. Among these clones, two apparent cell morphologies were discovered. Most of the clones grew adherently as multilayer and aggregates, we called them type 1 MHCC 97L (Fig. 4a), the other clone grew adherently as a uniform monolayer, and we called them type 2 MHCC 97L (Fig. 4b). Based on the statistics, the percentages of the type 1 and type 2 clones are respectively 97% and 3% (Fig. S7).

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