A Quantitative Perspective on Surface Marker Selection for the Isolation of Functional Tumor Cells

Cahall, Calvin F.; Lilly, Jacob L.; Hirschowitz, Edward A.; Berron, Brad J.

doi:10.4137/bcbcr.s25461

Cited by 16 publications

(22 citation statements)

References 65 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…From left to right: (A) Prior to culture DNA profiles are complex with a high proliferative/aneuploid fraction; (B) DAPI versus cytokeratin reveals that proliferating/aneuploid cells can be heterogeneous with respect to cytokeratin expression; (C) The fraction of cells coexpressing CD44 and CD90 is variable, ranging from 1. (23). Further supporting this data, HER-2/NEU (24) and EPCAM messenger RNA was present in MDA-MB-231 at low levels ( Fig.…”

Section: Resultssupporting

confidence: 74%

See 1 more Smart Citation

Antibody‐based cell‐surface proteome profiling of metastatic breast cancer primary explants and cell lines

et al. 2018

View full text Add to dashboard Cite

Flow cytometric cell surface proteomics provides a new and powerful tool to determine changes accompanying neoplastic transformation and invasion, providing clues to essential interactions with the microenvironment as well as leads for potential therapeutic targets. One of the most important advantages of flow cytometric cell surface proteomics is that it can be performed on living cells that can be sorted for further characterization and functional studies. Here, we document the surface proteome of clonogenic metastatic breast cancer (MBrCa) explants, which was strikingly similar to that of normal mesenchymal stromal cells (P = 0.017, associated with Pearson correlation coefficient) and transformed mammary epithelial cells (P = 0.022). Markers specifically upregulated on MBrCa included CD200 (Ox2), CD51/CD61 (Integrin α5/β3), CD26 (dipeptidyl peptidase-4), CD165 (c-Cbl), and CD54 (ICAM-1). Proteins progressively upregulated in a model of neoplastic transformation and invasion included CD26, CD63 (LAMP3), CD105 (Endoglin), CD107a (LAMP1), CD108 (Semaphorin 7A), CD109 (Integrin β4), CD151 (Raph blood group), and disialoganglioside G2. The proteome of the commonly used cell lines MDA-MB-231, MCF7, and BT-474 were uncorrelated with that of MBrCa (P = 1.0, 1.0, 0.9, respectively). The comparison has demonstrated the mesenchymal nature of clonogenic cells isolated by short-term culture of metastatic breast cancer, provided several leads for biomarkers and potential targets for anti-invasive therapy, including CD200, and highlighted the limitations of breast cancer cell lines for representing the cell surface biology of breast cancer. © 2017 International Society for Advancement of Cytometry.

show abstract

Section: Resultssupporting

confidence: 74%

“…The principal differences between MBrCa explants and epithelial breast cancer cell lines are a gain of mesenchymal markers (e.g., CD44, CD73, CD90, CD105) in MBrCa and (23). Further supporting this data, HER-2/NEU (24) and EPCAM messenger RNA was present in MDA-MB-231 at low levels ( Fig.…”

Section: Resultssupporting

confidence: 64%

Antibody‐based cell‐surface proteome profiling of metastatic breast cancer primary explants and cell lines

et al. 2018

View full text Add to dashboard Cite

show abstract

“…We have previously shown that A549 exhibit significantly lower surface density of epithelial markers (EpCAM and E-cadherin) than stem-like markers such as CD44 and metastasis markers like EGFR and ICAM-1. 33 Figure 5A shows that EpCAM targeting resulted in low photoinitiator loading density of below 1,200 eosin molecules per µm 2 . This is in good agreement with our previous findings as well as estimated EpCAM surface densities on tumor cell lines in the literature.…”

Section: Resultsmentioning

confidence: 99%

“…This is in good agreement with our previous findings as well as estimated EpCAM surface densities on tumor cell lines in the literature. 33, 34 The low eosin surface concentration using EpCAM resulted in low isolation yield near 1% for a PEGDA-575 monomer solution. For PEGDA-3500 in Figure 5B, eosin loading was also below 1,200 molecules per µm 2 and delivered correspondingly yield of intact cells at around 1%.…”

Section: Resultsmentioning

confidence: 99%

The Role of Surface Receptor Density in Surface-Initiated Polymerizations for Cancer Cell Isolation

Lilly

Berron

2016

Langmuir

Self Cite

View full text Add to dashboard Cite

Fluid biopsies potentially offer a minimally invasive alternative to traditional tissue biopsies for the continual monitoring of metastatic cancer. Current established technologies for isolating circulating tumor cells (CTCs) suffer from poor purity, yield, and require fixatives that preclude the collection of viable cells for longitudinal analyses of biological function. Antigen Specific Lysis (ASL) is a rapid, high-purity method of cell isolation based on targeted protective coatings on antigen-presenting cells and lysis depletion of unprotected antigen-negative cells. In ASL, photoinitiators are specifically labeled on cell surfaces that enable subsequent surface-initiated polymerization. Critically, the significant determinants of process yield have yet to be investigated for this emerging technology. In this work, we show that the labeling density of photoinitiators is strongly correlated with the yield of intact cells during ASL by flow cytometry analysis. Results suggest ASL is capable of delivering ~25% of targeted cells after isolation using traditional antibody labeling approaches.. Monomer formulations of two molecular weights of PEG-diacrylate (Mn~575 and 3500) are examined. The gelation response during ASL polymerization is also investigated via protein microarray analogs on planar glass. Finally, a density threshold of photoinitiator labeling required for protection during lysis is determined for both monomer formulations. These results indicate ASL is a promising technology for high yield CTC isolation for rare-cell function assays and fluid biopsies.

show abstract

“…The ability of conformal synthetic hydrogels to serve as optical labels, act as steric protective layers, and separate distinct cell types make these systems potentially useful for rare cell isolation and in vivo cellular therapies 10,11 .…”

Section: Introductionmentioning

confidence: 99%

Genetically Encoded Stimuli-Responsive Cytoprotective Hydrogel Capsules for Single Cells Provide Novel Genotype–Phenotype Linkage

Vanella

Bazin

et al. 2019

Chem. Mater.

View full text Add to dashboard Cite

Modification of cell surfaces with synthetic polymers is a promising approach for regulating cellular behavior. Here we describe a genetically controlled strategy for selectively encapsulating single yeast cells in synthetic microniches comprised of cross-linked phenol-modified alginate and chitosan hydrogel capsules. Our system links inducible gene expression with enzyme-mediated hydrogel polymerization and provides a novel genotype-phenotype linkage whereby only cells carrying a requisite gene encoding a flavin adenine dinucleotide (FAD)-dependent oxidoreductase undergo autonomous enzyme-mediated surface polymerization resulting in formation of hydrogel capsules. The composition of the hydrogel capsules is highly tunable and the capsule sizes are pH-responsive, allowing for control of capsule porosity and shell diameters over a range of 15-80 µm. The hydrogel capsules prevent extracellular proteins from reaching the cell surface, thereby conferring cellular immunity to lytic enzyme cocktails and rendering the hydrogel capsules cytoprotective against osmotic shock. We demonstrate the utility of this genetically controlled artificial hydrogel-encapsulated cell phenotype by isolating and enriching uniform eukaryotic cell lineages from genetically heterogeneous cell mixtures at 95-100% efficiency. The encapsulated cells remained viable and were capable of dividing and breaking free from their hydrogel capsules, allowing further propagation of selected cells. Our bottom-up approach to cellular compartmentalization links inducible intracellular genetic components with an artificial extracellular matrix that resists enzymatic lysis and mediates communication with the surrounding environment through a size-tunable and permeable hydrogel capsule.

show abstract

A Quantitative Perspective on Surface Marker Selection for the Isolation of Functional Tumor Cells

Cited by 16 publications

References 65 publications

Antibody‐based cell‐surface proteome profiling of metastatic breast cancer primary explants and cell lines

Antibody‐based cell‐surface proteome profiling of metastatic breast cancer primary explants and cell lines

The Role of Surface Receptor Density in Surface-Initiated Polymerizations for Cancer Cell Isolation

Genetically Encoded Stimuli-Responsive Cytoprotective Hydrogel Capsules for Single Cells Provide Novel Genotype–Phenotype Linkage

Contact Info

Product

Resources

About