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Isolation and Neuronal Reprogramming of Mouse Embryonic Fibroblasts
Juan M Adrian-Segarra1,2,3, Bettina Weigel1,2,3, Moritz Mall4,5,6
1Cell Fate Engineering and Disease Modeling Group, German Cancer Research Center (DKFZ) and DKFZ-ZMBH Alliance, Heidelberg, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|July 29, 2021
Summary
Researchers converted mouse embryonic fibroblasts (MEFs) into functional neurons using specific transcription factors. This direct neuronal reprogramming method offers a fast and efficient way to generate neurons for in vitro studies.
Area of Science:
- Neuroscience
- Cell Biology
- Molecular Biology
Background:
- Direct neuronal reprogramming converts non-neuronal cells into neurons.
- This technique is valuable for studying neurogenesis and generating neuronal models.
- Mouse embryonic fibroblasts (MEFs) are a common cell source for reprogramming.
Purpose of the Study:
- To provide a detailed protocol for direct neuronal reprogramming of MEFs.
- To generate functional mouse neurons in vitro using overexpression of key transcription factors.
- To enable versatile functional and mechanistic characterization of newly generated neurons.
Main Methods:
- Isolation of MEFs devoid of neural tissue.
- Overexpression of neuronal reprogramming factors (Ascl1, Brn2, and Myt1l) using lentiviral vectors.
- Direct conversion of MEFs into functional neurons.
Main Results:
- Successful direct reprogramming of MEFs into functional neurons.
- Efficient generation of mouse neurons in vitro.
- The protocol allows for quick and versatile characterization of reprogrammed neurons.
Conclusions:
- Direct neuronal reprogramming of MEFs is an effective method for generating functional neurons.
- This protocol facilitates in vitro studies of neurogenesis and neuronal function.
- The generated neurons can be used for diverse mechanistic and functional investigations.
