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Best practice for passaging murine embryonic enteric neuronal cell line before differentiation
Carmen D Rietdijk1, Lydia de Haan2, Richard J A van Wezel3,4
1Division of Pharmacology, Faculty of Science, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Universiteitsweg 99, 3584 CG, Utrecht, Netherlands.
Cytotechnology
|February 25, 2016
Summary
The immorto fetal enteric neuronal (IM-FEN) cell line is a model for studying enteric neurons. Optimal IM-FEN cell culture requires specific seeding density and confluency to manage unstable growth rates.
Area of Science:
- Neuroscience
- Gastroenterology
- Cell Biology
Background:
- The enteric nervous system (ENS) regulates gastrointestinal functions and is part of the gut-brain axis.
- The murine immorto fetal enteric neuronal (IM-FEN) cell line serves as a model for enteric neuron research.
- IM-FEN cells can be differentiated into neuronal phenotypes but do not produce action potentials in vitro.
Purpose of the Study:
- To optimize the culture conditions for the IM-FEN cell line.
- To identify factors influencing the growth rate of proliferating IM-FEN cells.
- To establish reliable protocols for using differentiated IM-FEN cells in experiments.
Main Methods:
- Culturing IM-FEN cells under varying conditions.
- Calculating a growth rate indicator based on seeding density and culture duration.
- Assessing differentiation success via neuronal protein staining.
- Analyzing the relationship between growth rate and culture parameters.
Main Results:
- Successful differentiation of IM-FEN cells into neuronal phenotypes was confirmed by positive protein staining.
- IM-FEN cell proliferation exhibited unstable growth rates.
- Growth rate was significantly influenced by seeding density and days in culture.
- Growth rate was independent of the culturist, prior overconfluency, or passage number.
Conclusions:
- Optimal IM-FEN cell culture involves maintaining 80% confluency before passaging.
- A seeding density of 0.0133 million cells/cm² is recommended for IM-FEN cultures.
- Researchers should anticipate unstable growth rates and potential overconfluency when culturing IM-FEN cells.
