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Cell surface antigen profiling using a novel type of antibody array immobilised to plasma ion-implanted polycarbonate
Heather Main1, Jelena Radenkovic, Elena Kosobrodova
1Department of Cell and Molecular Biology, Karolinska Institutet, 171 77, Stockholm, Sweden, heathermain187@gmail.com.
Insights
Researchers developed a novel cell capture microarray platform using plasma ion immersion implantation-treated polycarbonate (PIII-PC) to identify cell subpopulations based on surface antigen expression, offering a cost-effective method for biomedical research.
Area of Science:
- Biotechnology
- Cell Biology
- Immunology
Background:
- Identifying specific cell subpopulations is crucial for biomedical applications.
- Existing methods for cell discrimination can be costly and labor-intensive.
- There is a need for efficient and robust techniques to analyze cell surface antigen expressions.
Purpose of the Study:
- To develop a novel, cost- and labor-efficient array-based platform for discriminating cellular populations.
- To validate the platform's functionality and robustness using various cell types.
- To apply the platform to investigate neural stem cell differentiation and Notch signaling pathway effects.
Main Methods:
- Covalent immobilization of cluster of differentiation (CD) antibodies to plasma ion immersion implantation-treated polycarbonate (PIII-PC).
- Development of a transparent microarray matrix for direct light microscopy visualization.
- Validation of the PIII-PC array against flow cytometry, nitrocellulose arrays, and immunofluorescent staining.
Main Results:
- The PIII-PC array successfully discriminated cell populations based on unique surface antigen expression profiles.
- CD98 was identified as a novel marker for neural precursors.
- Polarized expression of CD9 was observed in neural rosettes, and CD9, CD49e, and CD117 were found in specific cell populations during Notch signaling modulation.
Conclusions:
- Covalent antibody array immobilization on PIII-PC surfaces provides accurate cell surface antigen data efficiently.
- This platform facilitates high-throughput identification and standardization of marker profiles in stem cell differentiation.
- The technology holds potential for applications in various genetic and disease contexts.
Abstract:
To identify and sort out subpopulations of cells from more complex and heterogeneous assemblies of cells is important for many biomedical applications, and the development of cost- and labour-efficient techniques to accomplish this is warranted. In this report, we have developed a novel array-based platform to discriminate cellular populations based on differences in cell surface antigen expressions. These cell capture microarrays were produced through covalent immobilisation of CD antibodies to plasma ion immersion implantation-treated polycarbonate (PIII-PC), which offers the advantage of a transparent matrix, allowing direct light microscopy visualisation of captured cells. The functionality of the PIII-PC array was validated using several cell types, resulting in unique surface antigen expression profiles. PIII-PC results were compatible with flow cytometry, nitrocellulose cell capture arrays and immunofluorescent staining, indicating that the technique is robust. We report on the use of this PIII-PC cluster of differentiation (CD) antibody array to gain new insights into neural differentiation of mouse embryonic stem (ES) cells and into the consequences of genetic targeting of the Notch signalling pathway, a key signalling mechanism for most cellular differentiation processes. Specifically, we identify CD98 as a novel marker for neural precursors and polarised expression of CD9 in the apical domain of ES cell-derived neural rosettes. We further identify expression of CD9 in hitherto uncharacterised non-neural cells and enrichment of CD49e- and CD117-positive cells in Notch signalling-deficient ES cell differentiations. In conclusion, this work demonstrates that covalent immobilisation of antibody arrays to the PIII-PC surface provides faithful cell surface antigen data in a cost- and labour-efficient manner. This may be used to facilitate high throughput identification and standardisation of more precise marker profiles during stem cell differentiation and in various genetic and disease contexts.
