Protocol for morphometric analysis of neurons derived from human pluripotent stem cells

Area of Science:

• Neuroscience
• Stem cell biology
• Developmental biology

Background:

• Analyzing neuronal morphology is crucial for understanding phenotypes and developmental changes.
• Human pluripotent stem cells (hPSCs) offer a valuable model for studying human neuronal development.
• Standardized protocols are needed for reliable morphometric analysis of hPSC-derived neurons.

Purpose of the Study:

• To describe an accessible protocol for the morphometric quantification of hPSC-derived neurons.
• To enable detailed analysis of neuronal morphology in 2D and 3D in vitro cultures.
• To provide a method for characterizing neuronal phenotypes during development.

Main Methods:

• Utilized free and accessible software tools for image analysis.
• Developed a protocol for morphometric quantification of neurons derived from hPSCs.
• Measured soma area, dendrite lengths (primary and secondary), and neurosphere area/perimeter.
• Applied the protocol to GFP-transfected neurons and immunostained neurospheres in vitro.

Main Results:

• Successfully quantified key morphological features of hPSC-derived neurons.
• Demonstrated the protocol's applicability to both 2D and 3D neuronal cultures.
• Provided measurements for soma area, dendrite lengths, and neurosphere characteristics.
• Established a reproducible method for neuronal morphometrics.

Conclusions:

• The described protocol offers a straightforward approach for neuronal morphometric analysis.
• This method facilitates the study of neuronal phenotypes and developmental trajectories using hPSC models.
• Accessible tools and techniques empower researchers to conduct detailed morphological investigations.