Biomarker discovery and cell enrichment

FITSAR methodology image for biomarker identification

Characterizing single cells allows for the possibility of identifying unique features corresponding to high-value populations. Our group seeks to discover truly novel biomarkers, with a concentration on mesenchymal stem cells. We have explored cellular characteristics spanning mechanophenotype, gene expression, and traditional surface proteins towards the goal of enriching function-specific stem/stromal cell types. These approaches have the potential to generate tailored stem cell populations that will perform a given function with a predictable baseline level, lessening the current problems with unstandardized cell sources used in regenerative medicine therapies.

  1. Dempsey ME, Chickering GR, González-Cruz RD, Fonseca VC, Darling EM. (2022) Discovery of surface biomarkers for cell mechanophenotype via an intracellular protein-based enrichment strategy. Cell Mol Life Sci. 79(320). PMCID: Pending. DOI: 10.1007/s00018-022-04351-w
  2. Dempsey ME, Woodford-Berry O, Darling EM. (2021) Quantification of antibody persistence for cell surface protein labeling. Cell Mol Bioeng. 14: 267-277. PMCID: PMC8175540. DOI: 10.1007/s12195-021-00670-3
  3. Sarnik SA, Sutermaster BA, Darling EM. (2020) Mass-Added Density Modulation for Sorting Cells Based on Differential Surface Protein Levels. Cytometry Part A. PMCID: PMC7855113. DOI: 10.1002/cyto.a.24192
  4. Sadick JS, Crawford LA, Cramer HC, Franck C, Liddelow SA, Darling EM. (2020) Generating Cell Type-Specific Protein Signatures from Non-symptomatic and Diseased Tissues. Annals of Biomedical Engineering. 4(8): 2218-2232. PMCID: PMC7416432. DOI: 10.1007/s10439-020-02507-y
  5. Sutermaster BA, Darling EM. (2019) Considerations for high-yield, high-throughput cell enrichment: fluorescence versus magnetic sorting. Sci Rep. 9: 227 (2019). PMCID: PMC6338736. DOI: 10.1038/s41598-018-36698-1
  6. González-Cruz RD, Sadick JS, Fonseca VC, Darling EM. (2018) Nuclear lamin protein C is linked to lineage-specific, whole-cell mechanical properties. Cell Mol Bioeng. 11(2): 131-142. PMCID: PMC5943047. DOI: 10.1007/s12195-018-0518-y PMCID: PMC5943047. DOI: 10.1007/s12195-018-0518-y
  7. Dempsey ME*, Marble HD*, Shin T, Fawzi NL, Darling EM. (2018) Synthesis and characterization of a magnetically active 19F molecular beacon. ACS Bioconjug Chem. 29(2): 335-342. PMCID: PMC5821531. DOI: 10.1021/acs.bioconjchem.7b00671 (*Co-first authors)
  8. Sadick JS, Darling EM. (2017) Processing fixed and stored adipose-derived stem cells for quantitative protein array assays. Biotechniques. 63(6): 275-280. PMCID: PMC5731247. DOI: 10.2144/000114620
  9. Sadick JS, Boutin ME, Hoffman-Kim D, Darling EM. (2016) Protein characterization of intracellular target-sorted, formalin-fixed cell subpopulations. Sci Rep. 6: 33999. PMCID: PMC5036045. DOI: 10.1038/srep33999
  10. Darling EM, DiCarlo D. 2015. High-throughput assessment of cellular mechanical properties. Ann Rev Biomed Eng. 17:35-62. PMCID: PMC8204286. DOI: 10.1146/annurev-bioeng-071114-040545Quartz crystal microplate
  11. Marble HD, Sutermaster BA, Kanthilal M, Fonseca VC, Darling EM. (2014). Gene expression-based enrichment of live cells from adipose tissue produces subpopulations with improved osteogenic potential. Stem Cell Res Ther. 5(5):145. PMCID: PMC4619280. DOI: 10.1186/scrt502
  12. Kanthilal M, Darling EM. (2014). Characterization of mechanical and regenerative properties of human, adipose stromal cells. Cell Mol Bioeng. 7 (4): 585-97. PMCID: PMC4255916. DOI: 10.1007/s12195-014-0350-y
  13. Desai HV, Voruganti IS, Jayasuriya C, Chen Q, Darling EM. (2014). Live-cell, temporal gene expression analysis of osteogenic differentiation in adipose-derived stem cells. Tissue Eng Pt A. 20(5-6): 899-907. PMCID: PMC3938923. DOI: 10.1089/ten.TEA.2013.0761
  14. González-Cruz RD, Darling EM. (2013). Adipose-derived stem cell fate is predicted by cellular mechanical properties. Adipocyte. 2 (2): 87-91. PMCID: PMC3661107. DOI: 10.4161/adip.23015
  15. González-Cruz RD, Fonseca VC, Darling EM. (2012). Cellular mechanical properties reflect differentiation potential of adipose-derived mesenchymal stem cells. Proc Natl Acad Sci USA. 109 (24): E1523-9. PMCID: PMC3386052. DOI: 10.1073/pnas.1120349109
  16. Darling EM, Guilak F. (2008) A neural network model for cell classification based on single-cell biomechanical properties. Tissue Eng Part A. 14 (9): 1507-1515. PMCID: PMC2748927. DOI: 10.1089/ten.tea.2008.0180