Limits...
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

Relationship between cell suspension concentrations and numbers of chamber containing one cell, none cell and more than one cell.White histograms refer to numbers of chamber with one cell, the gray histograms refer to numbers of empty chamber, the black histograms refer to chamber numbers with more than one cell.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4493670&req=5

f2: Relationship between cell suspension concentrations and numbers of chamber containing one cell, none cell and more than one cell.White histograms refer to numbers of chamber with one cell, the gray histograms refer to numbers of empty chamber, the black histograms refer to chamber numbers with more than one cell.

Mentions: In order to improve single cell isolation efficiency, series of condition optimizations were conducted. Firstly, we examined the effect of cell suspension concentration on single cell isolation efficiency, different concentrations such as 1.67 × 104, 3.00 × 104, 5.00 × 104, 1.42 × 105, 2.92 × 105, 5.33 × 105, 1.67 × 106 and 2.57 × 106 cells/mL were respectively examined as shown in Fig. 2. The results clearly demonstrated that the numbers of single cell chamber firstly increased with the increased cell suspension concentration, then decreased when the concentration reached to 2.92 × 105 cells/mL. The numbers of chamber without cell decreased with the increased cell suspension concentration during the whole range. Besides, the number of chambers included more than one cells was also increased with the increased cell suspension concentration. In consideration of the effect of the cell suspension concentration on the following single cell clonal cultivation, we here chose 3 × 105 cells/mL as the optimum concentration throughout the studies, about 598 lateral chambers included more than one cell under the optimized cell suspension concentration as showed in Fig. 2.


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)

Relationship between cell suspension concentrations and numbers of chamber containing one cell, none cell and more than one cell.White histograms refer to numbers of chamber with one cell, the gray histograms refer to numbers of empty chamber, the black histograms refer to chamber numbers with more than one cell.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Relationship between cell suspension concentrations and numbers of chamber containing one cell, none cell and more than one cell.White histograms refer to numbers of chamber with one cell, the gray histograms refer to numbers of empty chamber, the black histograms refer to chamber numbers with more than one cell.
Mentions: In order to improve single cell isolation efficiency, series of condition optimizations were conducted. Firstly, we examined the effect of cell suspension concentration on single cell isolation efficiency, different concentrations such as 1.67 × 104, 3.00 × 104, 5.00 × 104, 1.42 × 105, 2.92 × 105, 5.33 × 105, 1.67 × 106 and 2.57 × 106 cells/mL were respectively examined as shown in Fig. 2. The results clearly demonstrated that the numbers of single cell chamber firstly increased with the increased cell suspension concentration, then decreased when the concentration reached to 2.92 × 105 cells/mL. The numbers of chamber without cell decreased with the increased cell suspension concentration during the whole range. Besides, the number of chambers included more than one cells was also increased with the increased cell suspension concentration. In consideration of the effect of the cell suspension concentration on the following single cell clonal cultivation, we here chose 3 × 105 cells/mL as the optimum concentration throughout the studies, about 598 lateral chambers included more than one cell under the optimized cell suspension concentration as showed in Fig. 2.

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