Home › Analytical Chem › Cell-based Assay Protocol for the Prognostic Prediction of Idiopathic Scoliosis Using Cellular Dielectric Spectroscopy
Analytical Chem JoVE (Open Access) Citable · DOI

Cell-based Assay Protocol for the Prognostic Prediction of Idiopathic Scoliosis Using Cellular Dielectric Spectroscopy

DOI: 10.3791/50768-v
What you'll learn
  • ✓ Perform cellular dielectric spectroscopy (CDS) on peripheral blood mononuclear cells
  • ✓ Culture and treat PBMCs with recombinant osteopontin (OPN) protein
  • ✓ Interpret CDS results to predict idiopathic scoliosis prognosis
  • ✓ Complete a 4-day cell-based assay from fresh or frozen samples
Protocol

A structured protocol is presented for a cell-based assay as a functional test to predict the prognosis of idiopathic scoliosis using cellular dielectric spectroscopy (CDS). The assay can be performed with fresh or frozen peripheral blood mononuclear cells (PBMCs) and the procedure is completed within 4 days.

Difficulty
advanced
Total time
~4 days
Model organism
Human peripheral blood mononuclear cells (PBMCs)
Biosafety
BSL-1

Steps

1
Culture peripheral blood mononuclear cells

Prepare and culture PBMCs isolated from fresh or frozen peripheral blood samples. Establish appropriate culture conditions for downstream treatment and analysis.

▶ 02:04
2
Treat cells with recombinant osteopontin protein

Expose cultured PBMCs to recombinant OPN and incubate to induce cellular responses. This treatment primes cells for dielectric measurement.

▶ 03:38
3
Measure cellular dielectric spectroscopy and analyze results

Perform CDS measurements on treated cells to generate dielectric profiles. Analyze spectra to predict idiopathic scoliosis prognosis based on cellular responses.

▶ 05:49
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