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Related Concept Videos

Microtubules in Signaling01:22

Microtubules in Signaling

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The primary cilium, made up of microtubules, acts as antennae on the cell surfaces for relaying external stimuli into the cells. These fine hair-like structures are present, generally one per cell. These are non-motile cilia in a 9+0 microtubules arrangement, where the central pair of microtubules are absent. The primary cilia arise from the basal body embedded in the cell membrane. Intraflagellar transport (IFT) carries requisite proteins from the cytoplasm to the cilium because the primary...
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Related Experiment Video

Updated: May 6, 2026

Using Primary Neurosphere Cultures to Study Primary Cilia
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Analysis of Primary Cilium-Bearing Human Neuroprogenitors Using Flow Cytometry.

E De Gasperi1, M De Vita1, M Brusa1

  • 1Department of Molecular Medicine, University of Pavia, Pavia, Italy, unipv.eu.

Stem Cells International
|March 2, 2026
PubMed
Summary
This summary is machine-generated.

This study introduces flow cytometry to detect primary cilia on human neuroprogenitor cells, offering a scalable and objective method. This approach aids in studying ciliary defects linked to various diseases.

Keywords:
ciliopathyflow cytometryimmunostainingneuroprogenitorsprimary cilium

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Area of Science:

  • Cell Biology
  • Biotechnology
  • Medical Diagnostics

Background:

  • Primary cilia are crucial organelles involved in cell signaling, with defects linked to diseases like cancer and neurodevelopmental disorders.
  • Traditional methods for evaluating cilia, such as microscopy, are labor-intensive and operator-dependent.
  • Existing methods struggle with scalability and objectivity in assessing ciliary function.

Purpose of the Study:

  • To develop and validate a flow cytometry-based method for detecting and quantifying primary cilia on human neuroprogenitor cells (NPCs).
  • To establish a scalable, objective, and quantitative approach for assessing ciliary defects.
  • To demonstrate the feasibility of using flow cytometry for comparative analysis of ciliated cells in disease and control samples.

Main Methods:

  • Human induced pluripotent stem cells (iPSCs) were differentiated into NPCs.
  • NPCs were stained for ciliary markers (ARL13B, PERICENTRIN) and analyzed using flow cytometry.
  • Microscopy and imaging flow cytometry were used for validation of ciliary marker staining and localization.

Main Results:

  • Flow cytometry successfully detected and quantified primary cilia on human NPCs using specific ciliary markers.
  • The method confirmed the colocalization of axoneme and basal body markers on a single spot, indicative of intact cilia.
  • Flow cytometry effectively differentiated ciliary frequency between NPCs from ciliopathy patients and healthy controls.

Conclusions:

  • Flow cytometry provides a feasible, scalable, and operator-independent modality for detecting primary cilia on human NPCs.
  • This streamlined approach has discriminating capacity for studying ciliary defects in various pathological conditions.
  • The method offers a quantitative and objective alternative to traditional microscopy for ciliary analysis.